SPEX compositions and methods of use

ABSTRACT

The present invention provides novel polypeptides and other factors identified as being expressed in spleen tissue and related to lymphocyte activity and the immune response. These compounds are referred to herein as SPEX compounds. Human and murine homologues of SPEX polypeptides and polynucleotides are provided including two murine alleles. The invention further provides polynucleotides encoding the polypeptides and complementary nucleic acid sequences thereof. Also provided are immunogens, expression vectors, and antibodies related to the SPEX polypeptides and polynucleotides. The present invention discloses the use of anti-SPEX antibodies in the regulation of lymphocyte activity and the regulation of the immune response.

CROSS REFERENCE TO RELATED APPLICATION

[0001] The present application claims priority to U.S. ProvisionalApplication Ser. No. 60/467,206 filed Apr. 30, 2003, pending.

GOVERNMENT SUPPORT CLAUSE

[0002] This invention was made with government support under ContractAI44110 and AI31231, awarded by the National Institutes of Health. TheUnited States Government has certain rights in the invention.

UTILITY APPLICATION FOR U.S. PATENT

[0003] Filed Under 37 C.F.R. § 1.53(b)

BACKGROUND OF THE INVENTION

[0004] 1. Field of the Invention

[0005] The present invention is related to the fields of cellcommunication and signal transduction. In particular, this inventionprovides novel polypeptides and polynucleotides related to lymphocytedevelopment, proliferation, and cell linage commitment; immunogens;antibodies; vectors; host cells; and other compositions; and providesmethods of use thereof including for modulating lymphocyte activationand the immune response.

[0006] 2. State of the Art

[0007] Certain cell surface proteins are known to play critical roles inmodulating an immune response, including through cell to cell contactsand as receptors for soluble mediators. The identification andcharacterization of additional immune modulating proteins and methods ofmodulating lymphocyte development and activity remain important to thedevelopment of targeted therapies for a wide variety of conditionsincluding autoimmunity, cancer, transplant rejection, and inflammation.

[0008] The following polynucleotides are listed in the GenBank databasemaintained by the National Center for Biotechnology Information (NCBI),National Library of Medicine (NLM), Rockville Pike, Bethesda, Md. 20894:expressed sequence tags AA184189 and AA177302 and bacterial artificialchromosomes (BAC) pBeloBAC11 (Incyte Genomics, Palo Alto, Calif.),PBeloBAC.26056 (Incyte), and PBeloBAC.26057 (Incyte).

SUMMARY OF THE INVENTION

[0009] The present invention provides, in part, a novel gene encoding acell surface protein, referred to herein as SPEX. The name “SPEX” is acontraction of the first two letters in each word of the phrase “spleenexpressed” which is indicative of the high level of SPEX expressiondetected in spleen tissue. In certain embodiments, a substantially fulllength SPEX polypeptide is characterized as a signal transducingreceptor expressed on lymphocytes and capable of modulating lymphocyteproliferation, differentiation, development and/or cell lineagecommitment. In preferred embodiments, SPEX comprises a negativeregulator of an immune response. In one example, activation of a SPEXreceptor inhibits a metabolism of a lymphocyte or inhibits an immuneresponse. In another example, antagonism of a SPEX receptor enhances ametabolism of a lymphocyte or prolongs an immune response. Certainembodiments herein provide an antibody that immunoreacts with anextracellular domain of a SPEX polypeptide and a method of use forantagonizing the SPEX receptor, thereby stimulating lymphocytesexpressing the SPEX receptor.

[0010] In one embodiment, the present invention provides a SPEXpolypeptide comprising a multiple domain polypeptide including anextracellular domain, a transmembrane domain, and an intracellulardomain. The extracellular domain preferably includes a SPEXimmunoglobulin (Ig) like domain. The transmembrane domain preferablyanchors the SPEX polypeptide in a lipid bilayer, preferably acytoplasmic membrane of a cell and, alternately a liposome. The SPEXintracellular domain preferably includes one or more, and morepreferably three, tyrosine based domains.

[0011] Based, in part, on the disclosed domain structures, oneembodiment provides a SPEX polypeptide comprising a type I cell surfacesignal transduction phosphoprotein of the immunoglobulin superfamily.

[0012] The immunoglobulin superfamily is a multigene family thatincludes many cell surface proteins involved in molecular recognitionand adhesion. Ig like domains have a conserved core structure. Throughsequence analysis, the inventors identified an Ig like domain in theextracellular portion of the SPEX receptor molecule. Examples of apreferred SPEX polypeptide include an Ig like domain having a sequenceset forth in SEQ ID NO:3 of a human SPEX (hSPEX), SEQ ID NO:45 of amurine SPEX (mSPEX), or SEQ ID NO:88 of a second allele of mSPEX(referred to herein as mSPEXb). In one embodiment, a SPEX Ig like domainspecifically binds a dendritic cell (e.g., through specific binding ofthe SPEX polypeptide to a receptor/ligand on the dendritic cell).

[0013] Phosphoproteins are another multigene family. The phosphoproteinfamily includes many cell surface proteins involved in molecularrecognition and/or signal transduction. Phosphoproteins typicallyinclude a signaling domain that is integral to molecular recognition,phosphorylation, dephosphorylation, and signal transduction. CertainSPEX polypeptides comprise one or more tyrosine based domains. Inpreferred embodiments, the SPEX tyrosine based domain includes molecularrecognition, phosphorylation, dephosphorylation, and/or signaltransduction activities. For example, in one embodiment, a SPEXpolypeptide comprising a tyrosine based domain includes a tyrosineresidue capable of being phosphorylated and dephosphorylated. Examplesof preferred SPEX tyrosine based domains include SEQ ID NO:7, SEQ IDNO:9, and SEQ ID NO: 11.

[0014] Based in part on the Ig like and phosphoprotein structuresidentified in the SPEX polypeptide and/or data disclosed herein, certainembodiments provide that SPEX receptors modulate lymphocyte function.Certain embodiments provide a method of modulating a metabolism of alymphocyte including a SPEX receptor, comprising modulating an activityof the SPEX receptor. One embodiment provides a method of modulating aproliferation of lymphocytes that include a SPEX receptor, comprisingcontacting the lymphocytes with an anti-SPEX antibody that immunoreactswith an extracellular domain of the SPEX receptor. It is preferred thatthe proliferation of the lymphocytes, or the rate of proliferation, isincreased.

[0015] In certain embodiments it is contemplated that stimulating a SPEXactivity in lymphocytes that include a SPEX polypeptide will inhibit aproliferation of the lymphocytes (e.g., ligand binding of a SPEXreceptor contained in a lymphocyte).

[0016] In addition to an isolated polypeptide comprising an essentiallyfull length SPEX amino acid sequence, various domains thereof are usefulin methods of producing molecules that modulate SPEX activity. Forexample, a SPEX extracellular domain is useful to raise antibodies,which in turn are useful to modulate SPEX activity and lymphocytemetabolism; and a SPEX intracellular domain is useful to identify abinding partner, which in turn is useful to modulate SPEX phosphoproteinactivity and lymphocyte metabolism.

[0017] One embodiment of the present invention provides a SPEXpolynucleotide comprising a nucleic acid sequence encoding a SPEXpolypeptide, or a complement of the nucleic acid sequence. Furtherembodiments provide a nucleic acid for identifying a SPEX polypeptide, anucleic acid for amplifying a SPEX polynucleotide, and a SPEXpolynucleotide operably linked with a vector sequence. SPEXpolynucleotides are useful, for example, in methods of producing SPEXpolypeptides including in vitro and in vivo. A preferred cell forexpression of a SPEX polynucleotide comprises a lymphocyte including Bcells and T cells.

[0018] One embodiment provides a SPEX vector comprising a nucleic acidencoding a SPEX polypeptide, or a complement of the nucleic acid (a SPEXpolynucleotide); wherein the nucleic acid is operably linked to a vectorsequence. It is preferred that the vector sequence regulates ametabolism of the SPEX polynucleotide, and it is most preferred that thevector sequence regulates expression of the SPEX polynucleotide.

[0019] One embodiment provides a SPEX fusion sequence comprising a SPEXsequence operably linked to a heterologous sequence. Examples of certainuseful heterologous sequences include, but are not limited to: detectiontags, solubility enhancing factors, and biologically active factors.

[0020] One embodiment provides a SPEX variant sequence comprising a SPEXpolypeptide or a SPEX polynucleotide, wherein the SPEX sequence includesa modification or variation as disclosed herein. It is preferred thatthe SPEX variant sequence includes a structural and/or a functionalcharacteristic (i.e., an activity) of a non-modified SPEX sequence.

[0021] One embodiment provides a SPEX mutant sequence (polypeptide or anencoding polynucleotide). A preferred embodiment provides a polypeptidecomprising a SPEX amino acid sequence including a mutation of a tyrosinebased domain (e.g., substitution, deletion, truncation), morepreferably, a mutation of a tyrosine amino acid residue of the tyrosinebased domain. It is preferred that the mutation in a tyrosine baseddomain inhibits a SPEX signal transduction.

[0022] Another embodiment provides a host cell comprising a SPEX vector(a SPEX host cell). A SPEX host cell is useful, for example, in methodsof producing a SPEX polypeptide. In another example, a SPEX host cell isuseful in studies of lymphocyte metabolism and the modulation thereofincluding through the modulation of SPEX activation and/or inhibition.

[0023] One embodiment provides an isolated antibody, or fragmentthereof, that immunoreacts with a SPEX polypeptide of the presentinvention. In one embodiment, the anti-SPEX antibody modulates anactivity of an immune cell expressing a SPEX polypeptide. In a preferredembodiment, the anti-SPEX antibody inhibits a proliferation and/or adifferentiation of a mammalian cell expressing a SPEX polypeptide,preferably a lymphocyte, and most preferably a T cell, a B-cell, and/oran antigen presenting cell (APC).

[0024] One embodiment provides a method of screening candidate moleculesto identify a SPEX binding partner, comprising: contacting a candidatemolecule with a SPEX polypeptide; determining whether or not thecandidate molecule and the SPEX polypeptide form a binding complex,wherein the formation of a binding complex indicates that the candidatemolecule is a SPEX binding partner; and continuing the screening stepsuntil the SPEX binding partner is identified. Preferred candidatebinding partner molecules include small molecule organic compounds andpolypeptides. Preferred targets for the screening assay includes, but isnot limited to: a SPEX polypeptide; and more preferably a SPEXextracellular domain, a SPEX intracellular domain, a SPEX Ig likedomain, or a SPEX tyrosine based domain. One embodiment provides thatantagonists of a SPEX polypeptide or a SPEX domain activate a lymphocytecontaining SPEX receptors. Another embodiment provides that agonists ofa SPEX polypeptide or a SPEX domain inhibit activation of a lymphocytecontaining SPEX receptors.

[0025] One embodiment provides a method of purifying a SPEX polypeptidefrom a biological sample containing the SPEX polypeptide, comprising:contacting the biological sample with an affinity matrix having ananti-SPEX antibody attached to the matrix to produce an immunocomplexincluding the SPEX polypeptide and the antibody attached to the matrix;separating the remainder of the biological sample from the matrix;separating the SPEX polypeptide from the anti-SPEX antibody; andcollecting the SPEX polypeptide.

[0026] One embodiment provides a method of modulating a metabolism,preferably a proliferation, of a lymphocyte that expresses a SPEXreceptor polypeptide, comprising: contacting the lymphocyte thatexpresses the SPEX receptor polypeptide with an anti-SPEX antibody thatimmunoreacts with an extracellular domain of the SPEX receptorpolypeptide, thereby modulating the metabolism of the lymphocyte.Preferred lymphocytes include B cells and T cells.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The drawings form a portion of the specification of the presentinvention.

[0028]FIG. 1 provides a diagram of substantially full-length mouse andhuman SPEX polypeptides (mSPEX and hSPEX, respectively) havingextracellular, transmembrane (crosshatched), and intracellular domains(as labeled). The extracellular domains each include an immunoglobulin(Ig) like domain (horizontally hatched) and optionally have a cleavablesignal sequence (dashed areas), wherein the arrows point to the cleavagesite. The signal sequence is cleaved during post-translationalprocessing to generate the mature SPEX polypeptide. The intracellulardomains each include three tyrosine based motifs (vertically hatched).Each tyrosine based domain, in turn includes a tyrosine amino acidresidue (Y, is the one-letter code for tyrosine). The embodiments showndepict that the tyrosine can be optionally phosphorylated (encircled P).The relative position of the SPEX polypeptides in a biological membraneis shown. The stippled areas indicate portions of each polypeptidebetween the certain functional domains.

[0029]FIG. 2A provides a diagram showing embodiments of thecorrespondence between selected hSPEX polypeptides and the respectivesequence identifier for each selected polypeptide.

[0030]FIG. 2B provides a diagram showing embodiments of thecorrespondence between selected hSPEX polynucleotides and the respectivesequence identifier for each selected polynucleotide.

[0031]FIG. 2C provides a diagram showing certain embodiments of thecorrespondence between selected mSPEX polypeptides and the respectivesequence identifier for each selected polypeptide.

[0032]FIG. 2D provides a diagram showing certain embodiments of thecorrespondence between selected mSPEX polynucleotides and the respectivesequence identifier for each selected polynucleotide.

[0033]FIG. 3 provides four graphs (Panels 1-4) which demonstrate thatmonoclonal antibody PK18 inhibits T-cell activation. LN T cells werepurified from B10.BR and BALB.K mice and cultured in 96-well platescoated with mixtures of anti-CD3 and rat Ig (open symbols) or anti-CD3and PK18 (closed symbols). Data are expressed as the mean 3H-TdR(tritiated thymidine) incorporation of triplicate cultures (+/− standarddeviation) minus counts per minute (cpm) incorporation of cultures of Tcells alone (less than 1500 cpm). The incorporation of 3H-TdR into theDNA of the T cells is correlated with both proliferation of the T cellsand T cell activation.

DETAILED DESCRIPTION OF THE INVENTION

[0034] 1. SPEX

[0035] The present inventors identified, in part, a messenger RNA(mRNA), the expression of which is substantially increased bystimulating thymocytes with pharmacological activators used to model thein vivo thymocyte development program (i.e., thymocytes that arestimulated to proliferate and/or differentiate, see Example 1). Thepresent mRNA is referred to herein as “SPEX mRNA”. The inventors furtheridentified the gene, cDNA, and polypeptide corresponding to or encodedby the SPEX mRNA. Accordingly, the present SPEX polynucleotides and SPEXexpression products thereof are referred to herein, for example, as aSPEX gene, SPEX cDNA, SPEX mRNA, a SPEX antisense nucleic acid, and aSPEX polypeptide.

[0036] The present invention discloses that SPEX is expressed by both Tand B cells; with expression on B cells typically being greater thanexpression on T cells; activation of CD4 or CD8 T cells leads toup-regulation of SPEX expression in the T cells; activation of B cellsleads to down-regulation of SPEX expression in the B cells; SPEX isexpressed by CD4 and CD8 T cells, T cells that express the γδTCR (T cellreceptor), and CD25+CD4+ regulatory T cells; SPEX is induced in thethymus during the double positive to single positive transition; SPEX isupregulated during the pro B cell to pre B cell transition in the bonemarrow and then is further upregulated during the immature and mature Bcell developmental stages; SPEX is expressed on mature bonemarrow-derived dendritic cells; SPEX expression is low to absent onimmature bone marrow-derived dendritic cells; SPEX expression is low toabsent on NK (natural killer) cells; and SPEX is expressed on antigenpresenting cells in the spleen (including macrophages and dendriticcells).

[0037] In part, the present invention further provides compositions,such as: SPEX polypeptides and SPEX polynucleotides (including SPEXexpression vectors), antibodies that immunoreact with SPEX polypeptides,immunogens, and SPEX host cells. The present invention also providesmethods of using SPEX compositions, for example: to identify SPEXprotein binding partners; to isolate SPEX from natural or recombinantsources; to modulate lymphocyte metabolism; and/or as a marker orisolation tag to distinguish, identify, and/or purify lymphocytes thatexpress SPEX.

[0038] Definition of Purified

[0039] As used herein, the terms “isolated” and “purified” are usedinterchangeably and mean that the particular compound of interest isseparated from other contaminating substances so as to be free (i.e.,pure), or essentially free, from impurities including toxic matter suchas endotoxin (optionally discounting solutes, excipients, stabilizers,buffers, salts, pharmaceutically acceptable carriers, and the like whichare not necessarily contaminating substances). Accordingly, it ispreferred that a polypeptide, polynucleotide, immunogen, variant,mutant, fusion (polypeptide or polynucleotide fusion), vector, hostcell, antibody, and the like; of the present invention is in a purifiedform.

[0040] As used herein, when a purified compound of the present inventionis admixed or otherwise combined with a second purified compound orsubstance, the resulting composition is considered a “purified”composition (i.e., having defined components).

[0041] In light of the present invention, one skilled in the art canpurify a polypeptide of the present invention (including fragmentsthereof) using standard techniques for protein isolation andpurification. A pure polypeptide generally will yield a single majorband on a non-reducing polyacrylamide gel, subject to limited variation.For example, multiple bands closely spaced on an electrophoretic gel(e.g., a doublet or triplet of bands) often results from the presence ofa mixed population of post-translationally modified (or other) variantsof a polypeptide having a given amino acid sequence (e.g., a portion ofa population of a polypeptide is phosphorylated or otherwise modifiedwhile another portion of the population does not include themodification). Alternatively, polypeptide gene products of the allelesof a gene may differ in amino acid sequence (e.g., due to a geneticmutation). The present polypeptide gene products may migratedifferentially during electrophoresis, typically resulting in two bandscorresponding to the two alleles found in cells with the normalcomplement of chromosomal loci. Additional bands are also contemplated(e.g., due to differences in phosphorylation states and/or posttranslational protein modification such as glycosylation). Gel bands canbe examined to determine the content of the SPEX polypeptide in light ofthe present invention and general polypeptide sequencing techniques wellknown in the art such as sequencing (peptide or nucleotide) or massspectral analysis.

[0042] Such mixed populations of the polypeptide can be further purifiedto homogeneity (e.g., thereby providing a pure polyeptide), if desired(e.g., based on charge, size, affinity, or other methods known in theart). Also, the population can be treated to add a modification or toremove a modification to/from the polypeptides in a population (e.g.,phosphorylation can be added by a kinase or removed using a phosphatase,glycosylation can be removed by a glycosidase or added by a glycosyltransferase). The purity of the polypeptide can also be determined, forexample, by amino acid sequence analysis and/or by mass spectralanalysis.

[0043] In light of the present invention, one skilled in the art canpurify a polynucleotide of the present invention using standardtechniques for nucleic acid isolation and purification. For example,nucleic acids can be removed from proteins, lipids, and other compoundsusing standard nucleic acid preparation techniques. Mixed populations ofnucleic acid sequences can be resolved using agarose or polyacrylamidegel electrophoresis with collection of specific bands of interest.Nucleic acids can be identified and/or purified using sequence specificprobes. A sequence specific probe can be affixed to a matrix forconvenient isolation of a polynucleotide of interest through affinitychromatography. Polynucleotides can also be purified using cloningtechniques with selection of a single colony containing a singlepolynucleotide clone followed by isolation of the nucleic acid ofinterest from the host cell using standard techniques.

[0044] In light of the present invention, one skilled in the art canpurify an antibody of the present invention using standard techniques.Certain methods include affinity purification over an antigen boundmatrix (e.g., a SPEX polypeptide bound matrix) or general antibodypurification over protein A matrix.

[0045] In light of the present invention, one skilled in the art canpurify a host cell of the present invention using standard techniques.Certain methods include dilution plating techniques which may, or maynot, include use of a selectable marker gene contained in the host cell.

[0046] In certain embodiments, a composition including a purifiedpolypeptide, polynucleotide, antibody, host cell, or other compound ofthe present invention, may further include: buffer, water, salts,pharmaceutically acceptable carriers, adjuvants, fusions, labels, tags,markers, stabilizing agents, albumin, and the like, and/or proteinmodifications (e.g., phosphorylation or glycosylation). A purifiedcompound of the present invention may also include one or more of suchagents or be packaged with such agents and still be considered“purified”, as used herein (i.e., these agents are not necessarilycontaminating substances). It is preferred that the present compositionis a purified and sterile composition. For example, the sterilecomposition can be manufactured by combining a purified and sterilecompound of the present invention with one or more sterile buffer,conditioner, carrier, bulk, binding, or stabilizing agent under sterileconditions and/or by or sterilizing the composition after combining theSPEX compound and agent.

[0047] It is preferred that a purified and/or sterile polypeptide,polynucleotide, antibody or composition of the present invention issubstantially free of endotoxin or other pyrogens and undesirableirritants capable of inducing an adverse reaction when administered tohumans or other mammals. The present polypeptides, polynucleotides,antibodies, or compositions can be provided in dry form (e.g.,lyophilized), which form may include salts, stabilizers, etc as desired.It is preferred that the dry form embodiments are provided in acontainer, vessel, or vial capable of maintaining sterility and purityand, optionally, suitable for reconstitution with a reconstitution agent(see above) prior to administration to a human or other mammal. The SPEXcompound may be provided in a first container in a kit packaged togetherwith the reconstituting agent in a second container.

[0048] A SPEX Receptor

[0049] As used herein, a “substantially full length SPEX polypeptide”may be referred to as a “SPEX receptor”. It is preferred that the “SPEXreceptor” includes an activity as disclosed herein, including, but notlimited to: modulation of lymphocyte metabolism (e.g., activation,deactivation, or proliferation), cellular adhesion, phosphorylation,dephosphorylation (of a cytoplasmic domain). Preferred SPEX receptorsinclude an Ig like domain, a transmembrane domain, and a tyrosine baseddomain (preferably an intracellular domain). Highly preferred SPEXreceptors include, but are not limited to: SEQ ID NO:20, SEQ ID NO:21,SEQ ID NO:62, or SEQ ID NO:63. Alternative SPEX receptors include, butare not limited to SEQ ID NO:19 or SEQ ID NO:61. Additional alternativeSPEX receptors comprise SEQ ID NO:85 or SEQ ID NO:86 operably linked toa SPEX transmembrane domain and a SPEX intracellular domain.

[0050] 2. Polypeptides

[0051] Certain embodiments of the present invention provide a SPEXpolypeptide of mouse, human (preferred), or other mammalian origin. Anexample of a human SPEX (hSPEX) polypeptide comprises an amino acidsequence set forth in SEQ ID NO:20. An example of a murine SPEX (mSPEX)polypeptide comprises an amino acid sequence set forth in SEQ ID NO:62.One embodiment provides an allele of a murine SPEX, referred to hereinas mSPEXb. An exemplary sequence of the mSPEXb polypeptide comprises theamino acid sequence set forth in SEQ ID NO:86.

[0052] As used herein, the term “SPEX polypeptide” is meant to include asubstantially full length SPEX polypeptide, as well as, an amino acidsequence comprising (optionally, consisting essentially of) a domain,fragment, segment, fusion, mutant, and/or a variant thereof; asdescribed herein. It is preferred that a polypeptide of the presentinvention exist in a purified form.

[0053] A. Polypeptide Including A SPEX Immunoglobulin Like Domain

[0054] One embodiment of the present invention provides a polypeptidecomprising an amino acid sequence including a SPEX immunoglobulin (Ig)like domain. In certain embodiments, a SPEX Ig like domain is set forthin SEQ ID NO:3, SEQ ID NO:45, or SEQ ID NO:88.

[0055] In one embodiment, a preferred SPEX Ig like domain includes asecondary and/or tertiary structure, for example, an Ig fold or a betasandwich. In another embodiment, a SPEX Ig like domain maintains thesecondary and/or tertiary structure, such as the Ig fold or the betasandwich, when the SPEX Ig like domain is isolated or separated from apolypeptide sequence that normally contains the SPEX Ig like domain.

[0056] A SPEX Ig like domain is useful, for example, in the manufactureof an antibody that specifically binds SPEX and modulates signaltransduction, lymphocyte activation, lymphocyte development, or animmune response. In another example, a SPEX Ig like domain, optionally aSPEX extracellular domain, is useful to modulate a lymphocyte function,to modulate an immune response, or to screen for SPEX binding proteins.In still another example, a SPEX Ig like domain is useful in a processof identifying agonists and antagonists of lymphocyte and/or SPEXactivity.

[0057] Certain embodiments of a polypeptide comprising a SPEX Ig likedomain are set forth by sequence identifier in Table 1 below. Inembodiments that comprise a plurality of amino acid sequences that arenot necessarily continuous, it is preferred that the sequences areoperatively linked (e.g., by a peptide bond or by an amino acid sequencelinker). TABLE 1 CERTAIN EMBODIMENTS OF A POLYPEPTIDE INCLUDING AN IGLIKE DOMAIN A polypeptide including: Optionally further includingOptionally, not including (domain or SEQ ID NO) (domain or SEQ ID NO)(domain or SEQ ID NO) SPEX Ig like domain Transmembrane domain 3 5 SPEXIg like domain Tyrosine based domain 3 7, 9, or 11 3 7, 9, 11, 16, or 175 3 Transmembrane domain or 5 7, 9, or 11 12 or 13 Transmembrane domainor 5 5 12, 13, 14, or 15 16 or 17 19 or 20 1 21 45 47 45 7, 9, or 11 457, 9, 11, 58, or 59 5 45 Transmembrane domain or 47 7, 9, or 11 54 or 55Transmembrane domain or 47 47 54, 55, 56, or 57 58 or 59 61 or 62 43 6388 Transmembrane domain 88 Tyrosine based domain 85 or 86 Transmembranedomain 85 or 86 Transmembrane domain Tyrosine based domain 85, 86, or 88Transmembrane domain 85, 86, or 88 Transmembrane domain and tyrosinebased domain 85, 86, or 88 Transmembrane domain and 7, 9, and/or 11

[0058] Certain embodiments provide a polypeptide comprising(alternatively, consisting essentially of) an amino acid sequenceidentified in Table 1, above.

[0059] A method to determine whether a polypeptide of interest includesan Ig like domain comprises searching the Conserved Domain Databaseusing RPS-BLAST and inputting the amino acid sequence of interest as thesearch query. If the polypeptide includes an Ig like domain structure,then the position and sequence of the Ig like domain is specified by thesearch program along with alignments of Ig like domains from otherpolypeptides. Preferred search parameters are: search database, all;expect 0.01; filter, low complexity; search mode, multiple hits 1-pass.The Conserved Domain Database and Search Service v1.54 tool is providedby the Computational Biology Branch, National Center for BiotechnologyInformation (NCBI), National Library of Medicine (NLM), Rockville Pike,Bethesda, Md. 20894. Comparison (e.g., by sequence alignment) of thecandidate polypeptide to SEQ ID NO:3 or SEQ ID NO:45 is useful todetermine if the candidate polypeptide is a SPEX polypeptide, fragment,or variant as disclosed herein.

[0060] B. Polypeptide Including a SPEX Tyrosine Based Domain

[0061] Another aspect of the present invention provides a polypeptidecomprising a tyrosine based domain of a SPEX amino acid sequence. In oneembodiment, a tyrosine based domain comprises SEQ ID NO:7, SEQ ID NO:9,or SEQ ID NO: 11. A tyrosine based domain includes a tyrosine amino acidresidue. It is preferred that the tyrosine based domain comprises aphosphorylation site. It is also preferred that a SPEX tyrosine baseddomain is capable of being phosphorylated and desphosphorylated.

[0062] A SPEX tyrosine based domain is useful, for example, in a processof identifying a modulator of a SPEX signaling activity or lymphocyteactivation. For example, it is contemplated that a SPEX binding partnerthat specifically binds the intracellular domain of the SPEXpolypeptide, preferably at or near a tyrosine based domain motif, willinterfere with and thereby inhibit phosphorylation and/ordephosphorylation of the tyrosine based domain. This is expected tomodulate SPEX activity as the phosphorylation/dephosphorylation of thetyrosine based domain is contemplated to provide a pathway of SPEXsignal transduction. Accordingly, one embodiment provides a method ofmodulating a metabolism of a lymphocyte (alternatively a SPEX signaltransduction), comprising: administering a vector encoding a SPEXbinding partner to the lymphocyte wherein the binding partner binds anintracellular domain of the SPEX polypeptide, preferably a tyrosinebased domain. The binding partner vector expresses a polypeptide in thelymphocyte, wherein the polypeptide specifically binds the SPEXpolypeptide and modulates a lymphocyte metabolism and/or a SPEX signaltransduction.

[0063] Certain embodiments of a polypeptide comprising one or more SPEXtyrosine based domains are provided in Table 2, below. Table 2 alsoprovides alternative embodiments, wherein the polypeptide includes oneor more SPEX tyrosine based domains (left column), but does not includea second SPEX sequence or domain (right column). TABLE 2 CERTAINEMBODIMENTS OF A SPEX POLYPEPTIDE A polypeptide Optionally, thepolypeptide comprising: does not include SEQ ID NO: SEQ ID NO:  7 1, 2,3, 4, 5, 12, 13, 14, or 15  9 1, 2, 3, 4, 5, 12, 13, 14, or 15 11 1, 2,3, 4, 5, 12, 13, 14, or 15 7 and 9 1, 2, 3, 4, 5, 12, 13, 14, or 15 7and 11 1, 2, 3, 4, 5, 12, 13, 14, or 15 9 and 11 1, 2, 3, 4, 5, 12, 13,14, or 15 7, 9, and 11 1, 2, 3, 4, 5, 12, 13, 14, or 15 16 1, 2, 3, 4,5, 12, 13, 14, or 15 17 1, 2, 3, 4, 5, 12, 13, 14, or 15 18 1, 2, 3, 4,13, or 14 49 43, 44, 45, 46, 47, 54, 55, 56, or 57 51 43, 44, 45, 46,47, 54, 55, 56, or 57 53 43, 44, 45, 46, 47, 54, 55, 56, or 57 49 and 5143, 44, 45, 46, 47, 54, 55, 56, or 57 49 and 53 43, 44, 45, 46, 47, 54,55, 56, or 57 51 and 53 43, 44, 45, 46, 47, 54, 55, 56, or 57 49, 51,and 53 43, 44, 45, 46, 47, 54, 55, 56, or 57 58 43, 44, 45, 46, 47, 54,55, 56, or 57 59 43, 44, 45, 46, 47, 54, 55, 56, or 57 60 43, 44, 45,46, 54, or 55

[0064] Certain embodiments provide a polypeptide comprising(alternatively, consisting essentially of) an amino acid sequenceidentified in Table 2, above.

[0065] An immunoreceptor tyrosine based inhibitory motif (ITIM) is amotif known in the art to be included in certain immune cell signalingproteins, particularly immune cell membrane receptors. ITIMs participatein regulating immune responses, cell proliferation, clonal expansion,production of cytokines, cellular adhesion, and other biologicalactivities. An ITIM is set forth by IXYXXL (SEQ ID NO:93), wherein “X”represents any amino acid.

[0066] One embodiment of the present invention provides an ITIMcomprising the sequence IVYASL (SEQ ID NO:94). In one embodiment, thetyrosine based domain comprising GIVYASLNH (SEQ ID NO:9) includes theITIM set forth in SEQ ID NO:94. Another embodiment provides a SPEXpolypeptide comprising IVYASL (SEQ ID NO:94).

[0067] A signaling lymphocyte activation molecule (SLAM) associatedadapter protein (SAP) is another motif known in the art to be includedin certain immune cell signaling proteins. A SAP motif participates inregulating immune responses, cell proliferation, clonal expansion,production of cytokines, cellular adhesion, and other biologicalactivities. In one example, the SAP polypeptide, SH2D1A, is known in theart to inhibit signal transduction by SLAM so that the proliferation oflymphocytes such as T cells and natural killer cells does not continueunchecked. Defects in a SAP gene at Xq25 are associated with X-linkedlymphoproliferative disease (see, e.g., Buckley, R. (2000) NEJM343:1313-1324 and Sayos J. et al. (1998) Nature 395:462-469; eacharticle incorporated herein by reference).

[0068] One embodiment of the present invention provides a SAP bindingsite comprising the sequence TEYASI (SEQ ID NO:96). In one embodiment,the tyrosine based domain comprising EAPTEYASICVRS (SEQ ID NO: 11)includes the SAP set forth in SEQ ID NO:96. Another embodiment providesa SPEX polypeptide comprising TEYASI (SEQ ID NO:96).

[0069] C. SPEX Extracellular Domain

[0070] One embodiment of the present invention provides a polypeptidecomprising a SPEX extracellular domain or fragment thereof. In certainembodiments, the extracellular domain comprises SEQ ID NO:3, SEQ IDNO:45, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:54, SEQ ID NO:55, SEQ IDNO:85, SEQ ID NO:85, SEQ ID NO:86, or SEQ ID NO:88. In one embodiment,the extracellular domain further comprises a transmembrane domain (e.g.,SEQ ID NO:5 or SEQ ID NO:47). Alternatively other membrane anchoringsequences known in the art may be used. Numerous transmembrane sequencesare known that anchor polypeptides in plasma membrane, cell wall,cellular organelles, vesicles, liposomes, lipid rafts, and the like. Oneof ordinary skill in the art can use a transmembrane sequence tocombine, anchor, or otherwise operably link a polypeptide of the presentinvention to a one of the mentioned lipid based structures, preferably acellular plasma membrane.

[0071] D. SPEX Intracellular Domain

[0072] One embodiment of the present invention provides a polypeptidecomprising a SPEX intracellular domain or fragment thereof. In certainembodiments, the polypeptide comprises SEQ ID NO:7, SEQ ID NO:9, SEQ IDNO:11, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:58, or SEQ ID NO:59. Incertain embodiments, the intracellular domain further comprises atransmembrane domain (e.g., SEQ ID NO:5 or SEQ ID NO:47). In general,other membrane anchoring sequences can be used, if desired.

[0073] E. Immunogens

[0074] Certain embodiments of the present invention provide SPEXimmunogens. A “SPEX immunogen”, as used herein, is a SPEX polypeptide ora fragment thereof which is capable of eliciting an immune reaction. Inone embodiment, a SPEX polypeptide or fragment thereof is useful in themanufacture of an antibody that immunoreacts with a SPEX polypeptide orfragment thereof (see above). SPEX immunogens also are useful indiagnostic and experimental assays or kits. For example, certaindiagnostics and kits provide compositions and methods useful as a markerof development stage of lymphocytes, a marker of MAPK signaling pathwayactivation during the DP to SP stage of thymocyte development, a markerof cell type (cells that express SPEX are discussed above), or a markeror tag for purification and/or sorting of cell types based at least inpart on SPEX expression.

[0075] In general, a SPEX polypeptide including six or more consecutiveamino acid residues is capable of eliciting an immune response.Typically, increasing the number of amino acid residues in the SPEXpolypeptide enhances the immunogenic response to the peptide. Thus,certain embodiments provide, in increase order of preference, a SPEXimmunogen comprising 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, or 40 consecutive amino acids of the SPEX polypeptide.Alternatively, the SPEX immunogen comprises 6 to 9, 10 to 19, 20 to 29,30 to 39, or 40 to 50 consecutive amino acids of the SPEX polypeptide.

[0076] In certain preferred embodiments, a SPEX immunogen comprises aSPEX polypeptide domain, such as: an external domain, a transmembranedomain, an intracellular domain, an Ig like domain, a tyrosine baseddomain, or a signal sequence. Each SPEX polypeptide disclosed in Table 1and Table 2 is useful as a SPEX immunogen.

[0077] In preferred embodiments, a SPEX immunogen is purified. Forexample, purification of a SPEX immunogen from a longer SPEX polypeptideis preferred when the fragment is liberated from the longer SPEXpolypeptide using peptidases (e.g., tyrpsin and chymotrypsin) or otherpolypeptide cleaving agents (e.g., piperidine). One method formanufacture of a SPEX immunogen comprises solid phase polypeptidesynthesis, the general techniques of which are well known in the art.Another method comprises cellular expression and purification of a SPEXimmunogen.

[0078] The purified SPEX immunogen may be combined or linked with anycarrier and/or admixed with any adjuvant known in the art. The SPEXimmunogenic composition thereby created is still referred to herein as aSPEX immunogen. In regard to a purified SPEX immunogen, the adjuvant orcarrier is not necessarily considered a contaminant. Such carriers andadjuvants are useful to enhance the immunogenic response. Immunogenicityenhancing agents and their combination with polypeptides are well knownin the art. For example, Freund's complete adjuvant, or a carrierparticle, such as, keyhole limpet hemocyanin (KLH) or colloidal metals(e.g., colloidal gold).

[0079] F. Variants

[0080] Certain embodiments herein provide a polypeptide (referred toherein as a SPEX polypeptide variant) comprising an amino acid sequencehaving 95%, or more, sequence identity to a SPEX reference polypeptide.As used herein, a “SPEX reference polypeptide” is a polypeptide setforth by sequence identifier in Table 1 or Table 2. It is preferred thatthe present SPEX variant polypeptide comprises a purified SPEX variantpolypeptide. In general, the greater the sequence identity of the SPEXvariant polypeptide in comparison to the SPEX polypeptide, the morepreferred the variant polypeptide. Accordingly, certain embodimentsprovide, in order of increasing preference, a polypeptide comprising anamino acid sequence having 96%, 97%, 98%, or 99% sequence identity, ormore, to a SPEX polypeptide. A SPEX variant polypeptide herein includesat least one amino acid substitution, modification, addition, deletion,gap, and/or insertion when compared to a SPEX reference polypeptide.Preferred variants retain substantial biological activity compared to aSPEX reference polypeptide; accordingly, SPEX variant polypeptides areuseful, in general, in most embodiments wherein a SPEX referencepolypeptide is useful. The SPEX variant polypeptide is generallyconsidered herein as an alternative embodiment to a SPEX referencepolypeptide, however.

[0081] Sequence alignments and percent identity are determined in thepresent invention using JELLYFISH version 1.5 software by LabVelocity(LabVelocity, Inc., San Francisco, Calif.) using the followingparameters: ktuple size (1), number of top diagonals (5), window size(5), gap penalty (3), scoring method (percent), weight matrix (Gonnet),gap open penalty (10), gap extension penalty (0.2), residue specific gappenalties (yes), hydrophilic gap benefit (yes), gap separation distance(8), percent of identity for delay (30.0), output order (aligned). Anyor all of substitutions, deletions, insertions, additions, gaps, andinclusion of synthetic modified amino acid residues (e.g., non-naturallyoccurring amino acids) are meant to be included in the calculation ofpercent identity between two sequences. For example, substitution of anallo-isoleucine for an isoleucine or other amino acid residue in onesequence, would reduce the percent identity between sequences whenaligned. The inclusion of a D-amino acid in a sequence, wherein thecounterpart is an L-amino acid, will also reduce the percent identitybetween sequences.

[0082] Referring to a SPEX variant polypeptide including SEQ ID NO:3(alternatively, SEQ ID NO:48 or SEQ ID NO:88), it is preferred that thepolypeptide includes an Ig-like domain structure. For example, oneembodiment provides a variant polypeptide comprising an amino acidsequence that is 95% or more identical to SEQ ID NO:3 (alternatively,SEQ ID NO:45 or SEQ ID NO:88) and includes an immunoglobulin like domainstructure.

[0083] Procedures for making polypeptides with substitutions, deletions,insertions, additions, modified residues, gaps, etc. are routine in theart, and can be applied to the present polypeptides and variants inlight of the present disclosure. One method includes expression of apolypeptide from a polynucleotide encoding such changes (e.g., usingsite-directed mutagenesis to modify the polynucleotide), preferably withpurification of the expressed polypeptide. A preferred method ofproducing a variant polypeptide includes de novo synthesis of the SPEXvariant polypeptide, wherein the desired alteration(s) are made duringsynthesis.

[0084] It is preferred in certain embodiments that amino acidsubstitutions are conservative substitutions. In general, substitutionof a given amino acid in a wild type sequence with an amino acid havinga side chain with similar characteristics has a reduced impact on theresulting structure and function of the conservatively substitutedvariant polypeptide. Accordingly, certain embodiments provide aconservatively modified SPEX polypeptide including ten or fewer aminoacid changes (alternatively, no more than 10 amino acid changes),wherein the amino acid changes are conservative substitutions,additions, or deletions (preferably conservative amino acidsubstations). Fewer than 10 amino acid changes are preferred.Accordingly, in order of increasing preference, the conservativelymodified SPEX polypeptide includes 9 or fewer, 8 or fewer, 7 or fewer, 6or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1conservative amino acid change. Certain highly preferred conservativelymodified SPEX variants include SEQ ID NO:3, SEQ ID NO:45, or SEQ IDNO:88.

[0085] It is preferred that a SPEX polypeptide comprising SEQ ID NO:7,SEQ ID NO:9, or SEQ ID NO: 11 has no more than one conservative aminoacid substitution (alternatively an addition or a deletion) in SEQ IDNO:7, SEQ ID NO:9, or SEQ ID NO:11, respectively.

[0086] As used herein, the naturally occurring amino acids are groupedby their side chain characteristics for conservative substitution asfollows: aliphatic side chains (G, A, V, L, I); aliphatic side chainswith secondary amino group (P); aromatic side chains (F, Y, W); sulfurcontaining side chains (C and M, except wherein methionine is the firstamino acid of a polypeptide); aliphatic hydroxyl side chains (S, T);basic side chains (K, R, H); acidic side chains (D, E, N, Q). Forexample, alanine may be conservatively substituted for a valine presentin the non-variant (or wild type) polypeptide (the SPEX referencepolypeptide). Alternatively, valine may be conservatively substitutedfor an alanine present in the non-variant polypeptide. In anotherexample, proline does not have a conservative substitution. Althoughcysteine and methionine belong to a group of sulfur containing aminoacids, it is preferred that cysteine and methionine residues are notsubstituted (or deleted) in SPEX Ig like domain variant polypeptides. Itis also preferred that variant polypeptides including a SPEXimmunoglobulin like domain have a wild type complement of cysteineresidues within the Ig like domain to maintain disulfide formationand/or structural conformation of the domain.

[0087] G. SPEX Mutant Polypeptides

[0088] Certain embodiments provide a SPEX polypeptide including amutation (including a substitution, deletion, truncation etc.), whereinthe mutation inhibits a SPEX signal transduction and/or a modulation oflymphocyte metabolism. The present SPEX polypeptide is referred toherein as a “SPEX mutant polypeptide”. A SPEX mutant polypeptide differsfrom a SPEX polypeptide variant, for example, in that mutants having alow activity are preferred while variants having substantial activityare preferred. A “low activity” here is preferably an activity levelthat is 20% or less compared a reference SPEX polypeptide (preferably10% or less and more preferably 5% or less activity). The SPEX mutantpolypeptide is preferably purified.

[0089] A highly preferred SPEX mutant polypeptide comprises a mutationin a SPEX tyrosine based domain, even more preferably, the mutationcomprises a substitution or deletion mutation of a tyrosine residue ofthe tyrosine based domain. Preferably, the SPEX polypeptide includingthe tyrosine based domain mutant is characterized by an inhibited SPEXsignal transduction and/or a modulation of lymphocyte metabolism. A SPEXpolypeptide wherein the tyrosine based domains are not mutated (i.e.,wild type), preferably as set forth in SEQ ID NO:7, SEQ ID NO:9, and SEQID NO: 11 may be used for comparison of activity, etc.

[0090] Certain embodiments of a SPEX mutant polypeptide are set forth inTable 3 below. The numbering of the tyrosine residues of interest areprovided for exemplary hSPEX and mSPEX sequences, respectively. TABLE 3EXAMPLARY SPEX TYROSINE BASED DOMAIN MUTANTS Residue Number Relative tohSPEX SEQ ID NO: 21 226 257 282 Residue Number Relative to mSPEX SEQ IDNO: 63 245 274 299 SEQ ID NO: 21 Y Y Y Mutant #1 Y Y F Mutant #2 Y F YMutant #3 F Y Y Mutant #4 Y F F Mutant #5 F Y F Mutant #6 F F Y Mutant#7 F F F

[0091] Certain preferred SPEX mutant polypeptides comprise a dominantnegative SPEX mutant polypeptide, wherein the polypeptide inhibits theactivity of wild type SPEX polypeptides (e.g., by direct interaction,such as, multimer formation resulting in a defective multimer or byindirect action, such as, acting as a inhibitor for a ligand or anactivator of a wild type SPEX polypeptide). Each mutant #1 through #7 inTABLE 3 above is contemplated to comprise a dominant negative SPEXmutant polypeptide. SPEX mutant polypeptides are useful, for example, inmethods of modulating SPEX signaling a metabolism of a lymphocyte or animmune response. For example, the mutants may compete for a ligand of aSPEX receptor or act as a dominant negative inhibitor of SPEX activity.

[0092] H. Non-Naturally Occurring Polypeptide

[0093] In optional embodiments, a polypeptide of the present inventioncomprises a non-naturally occurring polypeptide. Examples ofnon-naturally occurring polypeptides include: (a) a polypeptide thatincludes an amino acid not found in nature or in the organism ofinterest, such as a synthetic amino acid or amino acid mimetic; (b) apolypeptide that includes a chemical moiety attached to the polypeptidethat is not associated with the polypeptide in nature, such as aradioactive isotope or a fluorescent marker; (c) a polypeptide thatcomprises a purified amino acid sequence wherein the full sequenceoccurs in nature, e.g., a purified truncated sequence; (d) a polypeptidethat does not include a portion of a naturally occurring polypeptide,such as an internal deletion; (e) a fusion polypeptide including two ormore amino acid sequences (the same or different sequences) joinedtogether, wherein the amino acid sequences are not found joined togetherin nature; (f) or a purified polypeptide removed from the milieu ofnaturally-occurring substances normally associated with the polypeptidein nature. The SPEX polypeptide may be purified, for example, from amammalian cell or from other cell types (e.g., bacterial, yeast, insect,plant) using techniques known in the art, including recombinant cloningin a host cell and purification therefrom. The polypeptide may bechemically synthesized, the techniques of which are well known in theart.

[0094] I. Polypeptide Fusions

[0095] One embodiment provides a polypeptide comprising at least a firstSPEX amino acid sequence operably linked to a second polypeptide,wherein the first and the second polypeptides are not found linked innature (referred to herein as a “SPEX polypeptide fusion). SPEX aminoacid sequences useful for making SPEX fusions are disclosed herein(e.g., the polypeptides disclosed in Table 1 and Table 2, the SPEXvariants disclosed herein, and the SPEX mutants disclosed herein). Theterms “SPEX fusion” and “SPEX fusion sequence” refer generically to SPEXfusion polypeptides and to SPEX fusion polynucleotides. A SPEX fusionpolypeptide of the present invention is isolated and preferably ispurified. In general, the polypeptide components that comprise a SPEXfusion can be linked in any desired order. SPEX fusion polypeptides areuseful, for example, to promote (enhance or increase) solubility,purification, expression, detection, selection, and antigenicity; of aSPEX polypeptide of interest.

[0096] One embodiment provides a polypeptide comprising a first SPEXpolypeptide operably linked to a heterologous polypeptide, wherein theheterologous polypeptide is different from the first SPEX polypeptide,and wherein the heterologous polypeptide and the first SPEX polypeptideare not normally found operably linked in nature. For example,“heterologous polypeptide” includes SPEX and non-SPEX sequences andsequences from the same or a different species.

[0097] In certain embodiments, the operable linkage is cleavable. Forexample, a SPEX fusion polypeptide may include a peptidase cleavage sitefor separating the SPEX polypeptide and the heterologous polypeptide.Cleavable peptide sequences are well known in the art and can beincorporated into a SPEX fusion polypeptide operable linkage (e.g., bycloning or de novo synthesis of the desired amino acid sequence), inlight of the present disclosure. Enzymes for such cleavage includetyrpsin, enterokinase, tissue plasminogen activator (tPA), factor Xa,furin, and others.

[0098] A preferred operable linkage of a SPEX fusion polypeptidecomprises a polypeptide bond (amide bond) joining each polypeptide ofthe fusion to one or more other polypeptides. Preferably, eachpolypeptide of a fusion is linked to another polypeptide of the fusionby a single polypeptide bond.

[0099] A preferred method for making a SPEX fusion polypeptide is tomake a contiguous polypeptide using genetic engineering techniques(i.e., expression of a polynucleotide engineered to combine the codingregions for the respective polypeptides by a peptide bond or amino acidlinking sequence). A SPEX fusion polypeptide can also be manufactured byde novo chemical synthesis (e.g., solid phase polypeptide synthesis).Optionally, the polypeptides of the fusion can be operably linked by anamino acid spacer sequence. Spacer sequences are useful, for instance,to separate functional domains in a fusion polypeptide (e.g., to avoidsteric hindrance). Such spacer sequences are known in the art andtypically including multiple glycine, alanine, and/or proline residues.

[0100] Alternatively, chemical cross linkers are also useful foroperably linking polypeptides. Chemical crosslinking of polypeptides isknown in the art and can be applied to making a SPEX fusion polypeptidein light of the present invention. Common reactive or functional groupsin chemical crosslinking reagents include: iuccinimidyl esters,maleimides, and iodoacetamides (kits for chemical crosslinking arecommercially available, for example, from Molecular Probes, Eugene,Oreg.). Common reactive groups of polypeptides include amino, carboxyl,sulfhydryl, aryl, hydroxyl and carbohydrates. Certain specific chemicalcrosslinking reagents include: DSP (Dithiobis(succinimidylpropionate)),DTSSP 3,3′-Dithiobis(sulfosuccinimidylpropionate), DSS (Disuccinimidylsuberate), BS³ (Bis(sulfosuccinimidyl) suberate), DST (Disuccinimidyltartrate), Sulfo-DST (Disulfosuccinimidyl tartrate), EDC(1-Ethyl-3-(3-Dimethylaminopropyl)carbodiimide Hydrochloride), DTME:Dithio-bis-maleimidoethane, SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate).

[0101] Other useful chemical crosslinkers include heterodifunctionalcrosslinkers which contain two or more different reactive groups andtypically allow for sequential conjugations with specific groups ofproteins, minimizing undesirable polymerization or self conjugation.Heterodifunctional crosslinkers which react with primary or secondaryamines include imidoesters and N-hydroxysuccinimide (NHS)-esters such assuccimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) andsuccimidyl-4-(p-maleimidophenyl)-butyrate (SMPB). Crosslinking reagentswhich react with sulfhydryl groups include maleimides, haloacetyls andpyridyl disulfides. Carbodiimide crosslinking reagents couple carboxylsto primary amines or hydrazides, resulting in formation of amide orhydrazone bonds. One widely used carbodiiumide crosslinking reagent is1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDAC),which is commonly used to couple carboxylic acids to amines and is anexample of a zero-length crosslinking reagent. These crosslinkingreagents are available from Pierce Chemical Co. (Rockford, Ill.) andSigma (St. Louis, Mo.), for example. Photoreactive crosslinking reagentsare also typically heterobifunctional crosslinking reagents. Upon UVillumination, these reagents react with nucleophiles or form C—Hinsertion products. Various methods for introducing reversible,substantially irreversible, and cleavable chemical crosslinks betweenpolypeptides are also well known in the art and can be used to make SPEXfusion polypeptides in light of the present disclosure.

[0102] A noncovalent interaction between two molecules that has veryslow dissociation kinetics can also function as an operable linkage. Forexample, avidin, streptavidin, NEUTRAVIDIN biotin-binding protein andCAPTAVIDIN biotin-binding protein can operably link up to four moleculesof a biotinylated target through high affinity, but non-covalentbinding.

[0103] In addition the heterologous sequence optionally comprises anydesirable biologically active factor, for example; an antibody, acytokine (e.g., interleukins, interferons, NF-κB, an IL-2R chain (Tacantigen)), a peptide hormone (e.g., EGF, TGFα, TGFβ), a chemokine, akinase, a phosphatase, a membrane translocation sequence or factor, annuclear translocation sequence, a membrane anchor, a toxin (e.g., ricin,the active portion of diphtheria toxin), a marker or identifying tag,and the like.

[0104] One embodiment provides a polypeptide comprising a SPEXpolypeptide operably linked to a heterologous polypeptide, wherein theheterologous polypeptide is substantially soluble in aqueous solution,preferably more soluble in aqueous solution than the SPEX polypeptide.Operably linking a SPEX polypeptide with a more soluble heterologouspolypeptide is desirable, for example, to assist in expression, bindingstudies, and in raising antibodies. A preferred substantially solubleheterologous polypeptide comprises a constant region of human IgG1, morepreferably the human IgG1 hinge CH2, and CH3 domains. In one embodiment,a solubility promoting heterologous polypeptide comprises SEQ ID NO:97.Another embodiment provides a polypeptide comprising a SPEX polypeptideset forth in SEQ ID NO:3, SEQ ID NO:45, SEQ ID NO:88, SEQ ID NO: 12, SEQID NO:54, or SEQ ID NO:86 operatively linked to a heterologouspolypeptide including a constant region of human IgG1; preferably thehuman IgG1 hinge, CH2, and CH3 domains. Protein A binds the constantregion of IgG1 with high affinity. Accordingly, a SPEX-human IgG1 fusioncan be purified using protein A affinity chromatography.

[0105] Examples of other contemplated solubility enhancing polypeptidesinclude thioredoxin (TRX), glutathione S-transferase (GST), Protein A,DsbA, and Escherichia coli maltose-binding protein (MBP) or solublefragments thereof. Another example includes the HAT epitope which is a19-amino-acid sequence from the chicken lactate dehydrogenase protein(BD Biosciences Clontech, Palo Alto, Calif.). The HAT sequence ofnon-adjacent histidine residues possesses less overall charge than tagswith consecutive His residues, such as the 6×His tag. As a result, HATpolypeptide fusions exhibit increased solubility compared to the 6×Histag fusions while still possessing strong affinity for immobilized metalions. The binding characteristics of the HAT sequence allow bothimidazole gradient and pH gradient purification of proteins under nativeconditions (e.g., at approximately neutral pH (pH 7), as well as underdenaturing conditions. An alternative embodiment provides a SPEXpolypeptide operably linked with a non-peptide organic solubilityfactor, preferably linked by a covalent bond to the SPEX polypeptide.

[0106] Examples of contemplated detectable labels or markers include:antibody recognized moieties, for example, FLAG epitope, DYKDDDDK (SEQID NO:101); c-myc epitope, EQKLISEEDL (SEQ ID NO:102), visualizationmarkers (e.g., green fluorescent protein, luciferase, biotin/avidin),enzymatic markers (e.g., horseradish peroxidase), radiolabels (e.g.,¹²⁵I, ¹⁴C, ³H, ³³P, ³⁵S, and ³²P), and combinations thereof (e.g.,radiolabeled and enzyme labeled antibodies).

[0107] Examples of contemplated purification tags or aids include: GST,MBP, TRX, calmodulin binding peptide (CBP), 6-His tag (or poly-His),FLAG, c-myc tag, radiolabels, fluorescent markers, and hemagglutinin(HA). These, and other moieties provide convenient purification of thecorresponding SPEX fusion on immobilized glutathione, maltose,phenylarsine oxide, calmodulin, and metal chelate resins, respectively.Such moieties are commonly utilized in commercially availablepurification systems. A fusion protein may also be engineered to containa proteolytic cleavage site located between the SPEX polypeptidesequence and the heterologous polypeptide sequence, so that a SPEXpolypeptide may be cleaved away from the heterologous moiety followingpurification. A variety of commercially available kits are available tofacilitate making a fusion polynucleotide including a desired insert,expressing a product from the insert, and purifying the product. Thesekits may provide vectors that place a cleavable linker between theproduct of interest and the heterologous polypeptide of the fusion suchthat the portions of the fusion product can be cleaved and purified(e.g., the CREATOR compatible expression systems, Clontech). In light ofthe present disclosure, these kits can be used to make a SPEX fusionsequence (polynucleotide and polypeptide).

[0108] J. Polypeptide Manufacture

[0109] In light of the present disclosure, a SPEX polypeptide of thisinvention can be made using a variety of techniques well known in theart for preparing or making a polypeptide. For example, by purificationfrom natural sources (e.g., lymphocytes or spleen or thymus tissue). Inanother example, a SPEX polypeptide can be made through production in ahost cell including in bacterial (e.g., K12), eukaryotic, yeast, insect,plant, mammalian, Chinese hamster (e.g., CHO), murine, and human cells(e.g., using transfer of a recombinant SPEX expression system). In stillanother example, SPEX polypeptide is made using synthetic de novomethods. In preferred embodiments, the SPEX polypeptide is purifiedusing methods known in the art for protein separation and purification,which methods are possible for isolating or purifying a SPEX polypeptidein light of the present invention. For example, in light of the presentinvention, one of ordinary skill in the art is able to use an anti-SPEXantibody disclosed herein to isolate or purify a SPEX polypeptideincluding specific fragments and domains thereof through affinityseparation.

[0110] Examples of useful amino acids (and nucleotides) are provided inthe World Intellectual Property Organization (WIPO) Handbook onIndustrial Property Information and Documentation, Standard ST.25:Standard for the Presentation of Nucleotide and Amino Acid SequenceListings in Patent Applications (1998), including Tables 1 through 6 inAppendix 2; hereinafter referred to as “WIPO Standard ST.25 of 1998”,incorporated herein by reference.

[0111] 3. Polynucleotides

[0112] One embodiment of the present invention provides a polynucleotide(referred to herein as a “SPEX polynucleotide”), comprising a nucleicacid sequence encoding a SPEX polypeptide, or a complement of thenucleic acid sequence. The complement of the nucleic acid sequence iscapable of hybridizing to the encoding nucleic acid sequence. It ispreferred that the SPEX polynucleotide comprises a purifiedpolynucleotide.

[0113] SPEX polynucleotides are useful, for example, in manufacturingSPEX polypeptides (e.g., in vitro or cellular polypeptide expressionwherein the SPEX polypeptide is preferably purified). SPEXpolynucleotides are also useful in methods that include administering aSPEX expression product to a cell, wherein the SPEX polynucleotideincludes an element for expression. A complement of the nucleic acidsequence encoding a SPEX polypeptide (a “SPEX complement”) is useful,for example, in assays or diagnostic kits suitable for detecting a SPEXexpression product. The present diagnostic kit, in turn, is useful todetect lymphocyte type or in methods of cell purification or sorting.

[0114] In one embodiment, the SPEX polynucleotide comprises a mammalianSPEX polynucleotide, preferably a murine SPEX (mSPEX) polynucleotide,and, more preferably, a human SPEX (hSPEX) polynucleotide. Oneembodiment provides a polynucleotide comprising a nucleic acid sequenceencoding a polypeptide set forth in Table 1 or Table 2 (or,alternatively, a SPEX variant polypeptide as disclosed above).

[0115] It is preferred that a polynucleotide of the present inventioncomprises a purified polynucleotide, which purified form issubstantially free of contaminating substances (discounting solutes,excipients, stabilizers, buffers, and the like) and removed from themilieu of substances with which SPEX occurs in nature. It is preferredthat a purified SPEX polynucleotide is substantially free of singlestranded oligonucleotide sequences used for polymerase chain reaction(PCR) amplification and/or sequencing.

[0116] Table 4 provides examples of correspondence between SPEX domainsand certain preferred polynucleotides along with polypeptides encoded bythe polynucleotides. The respective sequences are set forth by sequenceidentifier. TABLE 4 CERTAIN EMBODIMENTS OF SPEX POLYNUCLEOTIDES hSPEXmSPEX Nucleic Acid Amino Acid Nucleic Acid Amino Acid SPEXSequence/Domain SEQ ID NO SEQ ID NO SEQ ID NO SEQ ID NO Signal Sequence22 1 64/91 43/87 Ig Like Domain 24 3 66/92 45/88 Transmembrane 26 5 6847 Tyrosine based Domain 28 7 70  7 Tyrosine based Domain 30 9 72  9Tyrosine based Domain 32 11 74 11 Extracellular Domain (Processed) 33 1275/90 54/86 Extracellular Domain (Unprocessed) 34 13 76/89 55/85Extracellular (Processed) and 35 14 77 56 Transmembrane DomainsExtracellular (Unprocessed) and 36 15 78 57 Transmembrane DomainsTyrosine Based Region 37 16 79 58 Intracellular Domain 38 17 80 59Transmembrane & Intracellular 39 18 81 60 Domains Ig-Like Domain throughCOOH 40 19 82 61 SPEX (no Signal Sequence) 41 20 83 62 SPEX (includingSignal Sequence) 42 21 84 63

[0117] The embodiments in TABLE 4, above having more than one sequenceidentifier (e.g., “45/88” listed for a murine SPEX Ig like domain)refers to distinct specific alleles identified in the particularorganism.

[0118] As defined herein, the maximum length of a SPEX polynucleotidethat includes the adjacent genomic 5′ and 3′ regions with which a SPEXgene is normally associated in nature is 40,000 basepairs (discountingvector sequences, see below). To fall within the bounds of the presentinvention, a genomic clone of SPEX polynucleotide sequences mustnecessarily be no more than 40,000 basepairs (bases for single strandednucleic acids). It is preferred that the genomic clone, having 40,000basepairs or fewer, separated from other adjacent genomic sequencesnormally associated with a SPEX gene found in nature. As further definedherein, the minimum length of a SPEX polynucleotide is 18 residues(e.g., a polynucleotide consisting essentially of a nucleic acidencoding a polypeptide set forth in SEQ ID NO:94 or SEQ ID NO:96).

[0119] In preferred embodiments, a SPEX polynucleotide features apreferred length range. For example, one of ordinary skill in the art isaware that for each application or system using a polynucleotide, thereare typical length or length ranges that make efficient use of thepolynucleotide in the application. Accordingly one of ordinary skill inthe art is able to select a nucleic acid length best suited to anapplication of choice, in light of the present invention. Table 5 belowprovides examples of embodiments using a SPEX polynucleotide andpreferred lengths of SPEX polynucleotides for selected applications orsystems. TABLE 5 EXEMPLARY LENGTH RANGES OF SPEX POLYNUCLEOTIDESPreferred range for SPEX polynucleotide: (nucleotides or baseApplication or system of use: pairs as appropriate) SPEX polynucleotide18 to 40,000 (minimum to maximum) (discounting vector sequences) morepreferably 99 to 15,000 more preferably 150 to 7,000 most preferred 600to 2,000 SPEX insert for plasmid vector preferably 24 to 15,000 morepreferably 150 to 5,000 most preferred 630 to 1,500 SPEX insert forviral vector preferably 24 to 25,000 more preferably 180 to 10,000 mostpreferred 810 to 1,800 SPEX polynucleotide preferably 500 to 40,000insert in yeast artificial chromosome (YAC), more preferably 10,000 to35,000 bacterial artificial chromosome (BAC), still more preferably20,000 to 30,000 or cosmid

[0120] Referring to Table 5, above, length ranges are in nucleotideswhen referring to single stranded polynucleotides and basepairs whenreferring to double stranded nucleotides. The ranges of preferred SPEXpolynucleotide lengths disclosed in Table 5 are suggested for enhancedefficiency relative to each composition or application, but are notnecessarily absolute. In general, transformation efficiency of a givenvector system decreases with increasing size of the insert used in thevector.

[0121] One embodiment provides a polynucleotide, referred to herein as a“SPEX fusion polynucleotide”, comprising a nucleic acid sequence thatencodes a polypeptide including at least a first SPEX amino acidsequence operably linked to a second polypeptide, wherein the first andthe second polypeptides are not found linked in nature. For example, apolynucleotide comprising a nucleic acid sequence that encodes a SPEXfusion polypeptide. The present embodiment is especially useful forproducing a SPEX fusion polypeptide in quantity.

[0122] Certain preferred SPEX fusion polynucleotides include a nucleicacid sequence encoding one or more of a first amino acid sequence setforth by: SEQ ID NO:3, SEQ ID NO:45, SEQ ID NO:88, SEQ ID NO:12, SEQ IDNO:13, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:85, or SEQ ID NO:86,operatively linked to the a polypeptide set forth in SEQ ID NO:97. Thepresent fusion polynucleotides are useful in the production of anexternal domain of the SPEX polypeptide, wherein the fusion issubstantially soluble in aqueous solution, preferably more soluble thanthe SPEX polypeptide alone.

[0123] Compositions and methods are well known in the art for cleaving,hybridizing, and ligating segments of polynucleotide sequences, whichcompositions and methods are useful for the production of a SPEX fusionpolynucleotide in light of the present invention. If desired, a SPEXfusion polynucleotide may include a restriction site for separating thesegments of the SPEX fusion polynucleotide.

[0124] A. Intervening Sequence

[0125] In one embodiment a polynucleotide further comprises anintervening sequence (e.g., an intron) operably linked with a SPEXnucleic acid sequence. An intron operably linked with the SPEXpolynucleotide may be desirable, for example, to enhance transcriptionand to enhance stability of a mRNA expression product, particularly in amammalian expression system. Typically the intron is operably linkedbetween a first and a second segment of the SPEX nucleic acid sequence.Accordingly, one embodiment of the present invention provides apolynucleotide comprising a SPEX polynucleotide and an intron sequence,wherein the intron is operably linked to the nucleic acid upstream ofthe nucleic acid sequence. A preferred intron sequence is selected fromone or more SPEX genomic intron sequences. Optionally, the intronsequence is selected from one or more heterologous intron sequence or acombination of a SPEX intron and a heterologous intron.

[0126] Examples of useful intron sequences include a SV40 small-tantigen intron and an intron A from the CMV IE gene. These intronsequences are available commercially from Promega (Madison, Wis.) andInvivogen (San Diego, Calif.), respectively. It is preferred that thejunctions between the coding sequence and the intron sequence (i.e., thesplice junctions) comprise suitable splice sites for removal of theintronic sequence during processing (e.g., during a splicing reactionsuch as the processing of hnRNA to mRNA). In certain embodiments theSPEX polynucleotide includes any intron having suitable splicejunctions.

[0127] B. Degenerate Genetic Code Variant

[0128] One of ordinary skill in the art is aware of the degeneracy ofthe genetic code and is therefor able to use degenerate codons tomanufacture multiple nucleic acid sequences encoding the same SPEXpolypeptide in light of the present invention. For a table of thestandard genetic code and use thereof, as well as, the symbols forcommon amino acids and the abbreviations or codes thereof, seeBiochemistry, 3rd Edition, Stryer Ed., Freeman Publisher (1988) pages91-115 and inside back cover, incorporated herein by reference. Asuitable SPEX polynucleotide can be formulated, in light of the presentinvention, based upon the desired SPEX polypeptide sequence to beexpressed and knowledge of the standard genetic code.

[0129] C. Sequence Modification Techniques

[0130] Modifications to a SPEX sequence may be made, for example, duringchemical synthesis of the polymer (including polynucleotide andpolypeptide synthesis) or through mutagenesis (variegation) of apolynucleotide and, optionally, expression of an encoded polypeptidevariant. A preferred method of altering a polynucleotide sequence isthrough site directed mutagenesis. Methods for chemical synthesis,mutagenesis, site directed mutagenesis, and expression of apolynucleotide are well known in the art. Methods for confirmingproduction of the desired polynucleotide and polypeptide sequence arealso known in the art. These methods can be applied to making andconfirming SPEX sequence variants, in light of the present disclosureand knowledge in the art.

[0131] D. Vectors

[0132] In certain embodiments, it is desirable that a SPEXpolynucleotide further comprises a vector sequence. Accordingly, thepresent invention provides a purified polynucleotide, comprising avector sequence operatively linked to a nucleic acid sequence encoding aSPEX polypeptide, or a complementary sequence thereto. The nucleic acidencoding the SPEX polypeptide, or the complement thereof, are referredto herein as a “SPEX insert”. As used herein, a “SPEX vector” includes a“vector sequence” operably linked with a “SPEX insert”. SPEX vectorsequences are useful, for example, to express a SPEX mRNA, a SPEXpolypeptide, or a SPEX hybridizing sequence (e.g., an “antisense”nucleic acid). SPEX expression products may be labeled with a detectablelabel if desired. For example, the detectable label can be incorporatedduring synthesis. SPEX vectors are also useful as convenient cloningtools (e.g., shuttle vectors) or for production of SPEX nucleic acids incommercial amounts. Suitable SPEX inserts include SPEX polynucleotideslisted in the tables and sections herein, complements thereof, or encodeSPEX polypeptides disclosed herein (e.g., the SPEX polypeptidesdisclosed in Table 1 and Table 2, above).

[0133] A preferred use for a SPEX vector is in administering a SPEXexpression product to a cell. For example, a SPEX polypeptide can beadministered to a cell by introducing the SPEX vector into the cell,wherein the cell synthesizes the SPEX expression product using cellularmachinery. Expression of a SPEX polypeptide is useful, for example, tomodulate a metabolism of a lymphocyte, an immune response, and/or a SPEXsignal transduction.

[0134] As used herein, the SPEX insert may include untranslated regions,cloning sequences, intervening sequences, splicing sequences, or othersequences (limited by the disclosures above) in addition to the encodingregion. It is preferred that a SPEX vector is purified. A SPEX vectorincludes circular and linear nucleic acids.

[0135] The SPEX vector sequence preferably includes one or more controlelements for modulating the expression, replication, or other activityof the vector and/or the SPEX insert. Control elements include one ormore of a(n): affinity tag, branch point, cellular localization signal,enhancer, inducer, internal ribosome entry site (IRES), intron, Kozaksequence, polyadenylation site (poly A site), promoter, purificationtag, repressor, selectable marker, signal sequence, silencer, spliceacceptor, splice donor, start codon (initiator codon, translation startsite, ATG), stop codon, TATA box, terminator, and transcription startsite (e.g., Shine-Dalgarno sequence). The control elements may becontained in the vector sequence and/or the SPEX insert (optionally oneor more control elements can be located on another separate vector andact in cis). Numerous vector systems are available from commercial,academic, and other sources and can be readily operably linked with aSPEX insert using techniques well known in the art, in light of thepresent disclosure.

[0136] In certain preferred embodiments the SPEX vector is capable ofbeing expressed. Expression of a SPEX product from a SPEXpolynucleotide, preferably of a SPEX vector, includes: transcription ofan RNA from a DNA, translation of a polypeptide from an RNA or bothtranscription and translation. In one embodiment, expression comprisestranscription of the SPEX nucleic acid sequence encoding the SPEXpolypeptide forming a SPEX mRNA. It is preferred that the nucleic acidsequence encoding the SPEX polypeptide is “in frame” as contained in thevector. Most expression vectors are available, or easily adapted, forincorporation of an insert in the proper reading frame for synthesis ofthe desired polypeptide. A table of degenerate codons provides thepreferred correspondence between amino acid residues and the nucleotidecodon(s) that specify each amino acid residue during polypeptidesynthesis and is useful to predict or confirm with sequencing analysisthat a SPEX insert is in a desirable reading frame.

[0137] I. Promoters and Enhancers

[0138] It is preferred that a SPEX vector includes a promoter and/orenhancer for increasing the expression of the SPEX insert. In general,promoters and enhancers are control elements that modulate expression ofa transcript from a template nucleotide. The basic distinction betweenenhancers and promoters is operational and not absolute. Typically,promoter elements are located around the initiation site for RNApolymerase II (or other polymerase) and orient the direction oftranscription. Promoter elements usually include 7 to 20 bases ofnucleic acid and may contain one or more recognition sites fortranscriptional activators and/or repressors. Typically a module in eachpromoter functions to position the start site for RNA synthesis. A wellknown example of this is the TATA box. In general, enhancer elements arecapable of modulating (typically stimulating) transcription at adistance from the transcriptional start site. Enhancers may be locatedon the same nucleic acid as the start site of transcription (cis) or onanother nucleic acid (trans). In embodiments including a promoter, it isunderstood that the term “operably linked” means that the promoter is ina location and/or orientation in relation to the insert to control RNApolymerase initiation and expression of the transcript.

[0139] Promoter and/or enhancer elements may regulate the frequency oftranscriptional initiation. Typically, these are located in the region30-110 bp upstream of the transcriptional start site, although a numberof promoters are known to contain functional elements downstream of thestart site as well (e.g., within the transcribed nucleic acid). Thespacing between promoter elements is usually flexible, so that promoterfunction is preserved when elements are inverted or moved relative toone another. For example, in the tk promoter (from the herpes simplexvirus (HSV) thymidine kinase gene), the spacing between promoterelements can be increased up to 50 bp apart before activity begins todecline. Another example includes the lac operon. Depending on thepromoter, it appears that individual elements can function eithercooperatively or independently to activate transcription.

[0140] In certain embodiments, the particular promoter(s) that is(are)employed to control the expression of a SPEX insert is not believed tobe critical, so long as it is capable of expressing the polynucleotidein the targeted environment at desirable or sufficient levels for theintended embodiment (e.g., at a detectable level or over-expressedcompared to natively produced SPEX). Thus, for example, for expressionin a human cell, it is preferable to position the polynucleotide codingregion adjacent to and under the control of a promoter that is capableof being expressed in a human cell. Generally speaking, such a promotermight include either a human or viral promoter. Useful viral promotersinclude those from HSV tk, SV40 early transcription units,cytomegalovirus (CMV), mouse mammary tumor virus-long terminal repeat(MMTV-LTR), murine sarcoma virus (MSV-LTR), and Rous sarcoma virus(RSV-LTR). Useful promoters derived from human or other mammalian genesinclude those from β-actin and elongation factor 1-α (EF-1α). Numerousvectors are available that have promoter and enhancer elements suitablefor expression or modulated expression in a variety of environments.Certain preferred environments include human cells, preferablylymphocytes, and more preferably T-cells or B-cells.

[0141] By employing a promoter with well-known properties, the level andpattern of expression of a SPEX polynucleotide can be optimized. Forexample, selection of a promoter which is substantially more active inspecific cells, such as tyrosinase (melanoma), alpha-fetoprotein andalbumin (liver tumor), CC10 (lung tumor) and prostate-specific antigen(prostate tumor) will permit tissue specific expression of a SPEXpolynucleotide. Accordingly, in preferred embodiments, the promoterand/or vector is selected to drive transcription that is optimal in aparticular desired environment.

[0142] II. Non-Cellular Expression Environment

[0143] Non-cellular expression environments, commonly known as “in vitroexpression (systems)”, typically comprise cellular lysates or factorsfor transcription and/or translation of a template polynucleotidecontained in a reaction vessel. Preferably, the in vitro expressionsystem is at least partially refined to remove inhibitors and/orincrease reagent concentrations, and more preferably, substantiallypurified. In vitro expression systems are commercially available (e.g.,SP6 transcription kit, T7 transcription kit, SINGLE TUBE PROTEIN SYSTEM3, and RED NOVA LYSATE KIT each available from Novagen, Madison, Wis.and the RAPID TRANSLATION SYSTEM (RTS) available from Roche,Indianapolis, Ind.). A non-cellular expression system is preferred inthe event that a SPEX expression product of interest is toxic to thehost cell in a cellular expression environment. One embodiment providesa SPEX expression vector adapted for use in an in vitro expressionsystem.

[0144] III. SPEX Host Cells

[0145] Useful cellular expression environments include prokaryotic andeukaryotic host cells. The term “host cell” refers to a cell including aSPEX polynucleotide, preferably a SPEX vector, wherein the SPEXpolynucleotide is introduced to the cell through the hand of man to forma SPEX host cell (e.g., a recombinant, non-naturally occurring, SPEXpolynucleotide transferred into a cell for the purpose of expression).Accordingly, one embodiment provides a host cell including a SPEXvector. The present “host cell” is referred to herein as a “SPEX hostcell”. Preferably, the SPEX vector comprises a SPEX expression vectorand preferably is capable of producing a SPEX expression product.

[0146] In preferred embodiments, the SPEX host cell does not have SPEXexpression in the absence of a transformed SPEX polynucleotide suitablefor expression (e.g., a recombinant SPEX expression vector). Cells thatexpress SPEX without the introduction of a SPEX polynucleotide by thehand of man have a “native SPEX expression” as used herein. SPEXexpression from a SPEX polynucleotide transformed into the cell by thehand of man refers herein to “an exogenous SPEX expression” or a“recombinant SPEX expression”. Exogenous SPEX expression is meant toinclude SPEX expression from a SPEX polynucleotide incorporated into thegenome of the host cell in a process directed by the hand of man. Inoptional embodiments, the SPEX host cell may have a native SPEXexpression; however, it is preferred that the expression product of theexogenous SPEX polynucleotide is in greater abundance than the nativeSPEX product (i.e., it is preferred that the recombinant SPEXpolynucleotide is over expressed).

[0147] Certain embodiments provide a polynucleotide including a vectorsequence operably linked with a SPEX insert, wherein the vector sequenceincludes one or more control elements for expression of the SPEX insertin a host cell; preferably one or more of a bacteria cell, a yeast cell,an insect cell, or a mammalian cell. Numerous vectors and host cellsuseful for expression of the SPEX insert in light of the presentinvention are available from academic, commercial, and other sources.Certain additional examples of useful host cells include: Escherichiacoli (E. coli) and the K12 strain of E. coli (bacterial cells); theyeast cells Schizosaccharomyces cerevisiae (S. cerevisiae), S. pombe,and Pichia pastoris (P. pastoris); the insect cells Spodopterafrugiperda 9 (Sf9), Sf21, Trichoplusia ni (commonly referred to as HighFive cells), and S2 (drosophilae); and the mammalian cells includeHEK-293 (human kidney), Chinese hamster ovary (CHO), Bowes Melanoma(human melanoma), HeLa (ovarian carcinoma), CHU2 (human oral tumor), andHBEC-90 (human brain endothelium), 33.1.1 (mouse pre-B cell), K46 (mouseB lymphoma), TR2 (mouse oligodendritic). Certain preferred mammaliancells include: lymphocytes, T-cells, B-cells, immature T-cells, immatureB-cells, differentiated T-cells, differentiated B-cells, and naturalkiller cells. These cells, and others, are available from e.g.,Invitrogen, Novagen, CLONTECH, and the American Type Culture Collection(ATCC, Manassas, Va.).

[0148] In certain embodiments, it is desirable to express the SPEXinsert in more than one host cell. Accordingly, in certain embodimentsit is preferred to use a vector system having control elements suitablefor expression of the SPEX insert in multiple host cell types. Forexample, the TRIEX MULTISYSTEM EXPRESSION VECTORS (CALBIOCHEM, La Jolla,Calif.) includes optimized transcription and translation signals forexpression in E. coli, baculovirus, and mammalian cell types.

[0149] Expression in Bacteria

[0150] Useful bacterial cell cloning vectors for cloning with a SPEXinsert include: pUC8, pUC9, pBR322, pBR329, pBC-SK, pBC-KS, LAMBDA ZAPII (available from academic sources, Promega (Madison, Wis.), orStratagene (La Jolla, Calif.). It is preferred that host cells includean origin of replication capable of functioning in the given cellularenvironment. For example, bacterial vectors may include a E. coli originof replication (ori).

[0151] The pET vector series (Novagen (Madison, Wis.), Promega,Stratagene) contains the T7 promoter and the T7 gene 10 translationinitiation signals useful for driving high level expression of a SPEXinsert in bacterial cells that include the T7 factors. The pET vectorsystem is made inducible by transfection into bacterial cells that lackT7 RNA polymerase, for example, BL21 (DE3). Expression is induced bytransfection with a vector for expressing the missing T7 polymerase orby removing repression of native T7 expression.

[0152] The pL Expression System (Invitrogen, Carlsbad, Calif.) alsoprovides tight inducible transcriptional control of bacterial expressionsystems. The pL vectors include the strong pL promoter and are capableof driving expression of a SPEX insert. The pL promoter is controlled bythe lambda cI repressor protein which is expressed in the E. coli host.The cI repressor gene is engineered into the bacterial chromosome underthe control of the tightly-regulated trp promoter. Expression of theSPEX polynucleotide can be induced by the addition of tryptophan.

[0153] The pBAD vectors (Invitrogen) are useful for inducible expressionof an operably linked SPEX insert driven from the araBAD promoter whichcan be modulated to stimulate or repress transcription of the SPEXinsert by the addition of arabinose or glucose, respectively, to theculture medium. Additional vectors that include one or more controlelements for driving expression in bacterial systems include: Trc/Tacpromoter vectors (CLONTECH, Palo Alto, Calif.), Lambda PR promotervectors (Pharmacia, Peapack, N.J.), and Phage T5 promoter based vectors(QIAGEN, Valencia, Calif.). Embodiments provide a SPEX insert operablylinked with each vector or any vector, in general, including one or morecontrol elements for expression a product of a SPEX insert in abacterial cell.

[0154] Expression in Yeast

[0155] Certain embodiments provide a polynucleotide including a vectorsequence operably linked with a SPEX insert, wherein the vector sequenceincludes one or more control elements for expression of the SPEX insertin a yeast cell. It is preferred that a SPEX expression vector capableof expression in yeast includes a yeast origin of replication (e.g.,ColE1).

[0156] The pESC vectors (Stratagene) include GAL1 and GAL10 controlelements for expression in yeast cells, preferably S. cerevisiae. ThepESC vector set includes expression one or more different expressionproducts using selectable features including: a FLAG epitope; a c-mycepitope (tag); and HIS3, TRP1, LEU2, or URA3 selectable markers.

[0157] The pYES vector set (Invitrogen) includes a GAL1 promoter and isuseful for expressing a SPEX insert in yeast cells, preferably S.cerevisiae. The ESP Yeast Protein Expression and Purification System(Invitrogen) includes a nmtl promoter of S. pombe and is useful forexpressing a SPEX insert in yeast cells, preferably S. pombe. The ESPvector set includes expression one or more different expression productsusing selectable features including: a FLAG epitope; a c-myc epitope(tag); and HIS3, TRP1, LEU2, or URA3 selectable markers. Certain ESPvectors further include a glutathione s transferase (GST) peptide tagfor convenient purification of yeast expressed SPEX-GST fusionpolypeptides by GST affinity chromatography. The SpECTRA S. pombeExpression System (Invitrogen) includes additional vectors useful forconstruction of a SPEX vector capable of expression in yeast. The Pichiapastoris Expression System (Invitrogen) includes vectors useful forconstruction of a SPEX expression vector capable of expression in yeast,preferably P. pastoris, and bacteria, preferably E. coli.

[0158] Expression in Insect Cells

[0159] Certain embodiments provide a polynucleotide including a vectorsequence operably linked with a SPEX insert, wherein the vector sequenceincludes one or more control elements for expression of the SPEX insertin an insect cell.

[0160] The DES (Drosophila Expression System, Invitrogen) is optimizedfor expression of a SPEX insert expression in Drosophila cells,preferably S2 cells. The DES vectors pAc5.1/V5-His and pMT/V5-Hisprovide for expression promoted by an Ac5.1 or V5 promoter, respectivelyand include a His tag for detection and/or purification. ThepMT/BiP/V5-His vector further provides the BiP Drosophila secretionsignal sequence for secretion of an expressed polypeptide.

[0161] The INSECTSELECT System (Invitrogen) is useful for expression ofa SPEX insert in a broad range of insect cell lines including Sf9, Sf21,HIGH FIVE, and S2; preferably for the expression of secreted proteins.

[0162] Expression in Mammalian Cells

[0163] Certain embodiments provide a polynucleotide including a vectorsequence operably linked with a SPEX insert, wherein the vector sequenceincludes one or more control elements for expression of the SPEX insertin a mammalian cell, preferably a human cell.

[0164] Adenoviral and retroviral vectors are useful for expressing aSPEX insert in mammalian cells. The ViraPower Adenoviral ExpressionSystem, for example, includes E1 and E3 deleted, pAd-DEST adenovirusbased vectors having a human CMV expression promoter or without promoterfor convenient insertion of a promoter of choice. Packaging of theadenovirus particles is described in the manufactures literature(Invitrogen). The ViraPower Lentiviral Expression System, in anotherexample, is optimized for stable transduction of dividing andnon-dividing cells (Invitrogen).

[0165] The pShooter vectors (Invitrogen) include EF-1α or CMV promotersand may further include a signal sequence capable expressing a SPEXpolypeptide and of directing the expressed SPEX polypeptide to aspecific cellular compartment. For example, the pEF/myc/cyto vectorincludes an EF-1α promoter and a c-myc tag. A signal sequence is notincluded with the present vector for obtaining cytoplasmic expression.The pEF/myc/nuc vector includes an EF-1α promoter, a c-myc tag, and anuclear localization sequence (from SV40) capable of directing anexpressed SPEX polypeptide to the nucleus of a mammalian cell. ThepEF/myc/mito vector includes an EF-1α promoter, a c-myc tag, and amitochondrial localization sequence (from COX VIII cDNA) capable ofdirecting an expressed SPEX polypeptide to the nucleus of a mammaliancell. Another set of pShooter vectors include a CMV promoter whichreplaces the EF-1α promoter. The pShooter vectors are capable of drivingexpression in a broad range of mammalian cell types include most humancells and tissues.

[0166] The pSG5 vector (Stratagene) is capable of driving expression ofa SPEX insert in a broad range of cell types including mammalian(including human) and bacterial cell types. Mammalian expression isdriven by an SV40 early promoter and bacterial expression is driven by aT7 promoter. The pSG5 vector preferably further includes an ori,ampicillin resistance gene, and a phage f1 origin (e.g., allows rescueof ssDNA for use in mutagenesis and sequencing).

[0167] The pIRES-hrGFP-1 vector (Stratagene) is capable of expressing aSPEX insert and a marker gene (in this example, a humanized greenfluorescent protein (hrGFP)) using a CMV promoter. The vector includesan internal ribosome entry site (IRES) providing for expression of boththe SPEX insert and the marker gene from a single expressed mRNAspecies. Thus, the SPEX and the marker gene are expressed as a fusionmRNA and expressed as separate polypeptides. Other SPEX expressionvectors may further include an IRES, if desired.

[0168] In certain embodiments, the SPEX vector construct furtherincludes a segment encoding another expression product. For instance,the vector may include a selectable marker, detectable label, and/orpurification label. Examples include a segment encoding: amp, neo,hygro, zeo, puro, mycophenolic, kan, enhanced green fluorescent protein(EGFP), enhanced cyan fluorescent protein (ECFP), enhanced yellowfluorescent protein (EYFP), DsRed2, HcRed1, myc tag, FLAG tag, and GSTtag. In certain embodiments, the vector is selected to provide the labelpositioned to form an operably linked fusion with the SPEX insert.Certain vectors further provide an internal ribosome entry site (IRES).See, e.g., U.S. Pat. No. 4,937,190 to Palmenberg et al., incorporatedherein by reference. In one embodiment, an IRES facilitates theexpression of two proteins (at least one comprising the SPEXpolypeptide) in animal cells using a single-transcript vector (STV).

[0169] Cellular expression of SPEX typically involves introduction ofthe SPEX expression construct into a cell of choice wherein cellularfactors drive expression. Method for introducing expression vectors intoa cell are known in the art (e.g., calcium phosphate precipitation,lipid based transfection, peptide based membrane translocation,electroporation, and the like) See U.S. Pat. No. 6,312,956 to Lane andU.S. Pat. No. 6,165,720 to Felgner et al., each patent incorporatedherein by reference in its entirety.

[0170] Table 7 below provides preferred embodiments of SPEX vectors andTable 6 provides preferred features and uses for these SPEX vectorconstructs. TABLE 6 PREFERRED SPEX EXPRESSION VECTORS Vector BackboneDescription SPEX Vector Size Backbone RE sites Designation (kb) SPEXInsert Designation Backbone Source Insert/Backbone pSPEX-YFP 5.7 957 bpSPEX gene pEYEP-N1 CLONTECH Bgl II/Bam H1 (SEQ ID NO: 106) SPEXYFP- 6.81.7 bp fragment of pBABEpuro Nucleic Acids Not1 fill Bgl II/ BABEPUROpSPEX-YFP Res. (1990) SnaBI, BamHI (SEQ ID NO: 107) 18: 3587-3596pSPEX-Bgl II 5.4 1276 bp SPEX gene pCR 2.1 Invitrogen Bgl II/Bam H1 (SEQID NO: 109) TOPO pSPEX-Bgl II- 6.4 1276 bp SPEX gene pBABEpuroInvitrogen Bgl II/Bam H1 BABEpuro (SEQ ID NO: 109) pSPEXCAL 6.7 957 bpSPEX gene pCAL-C Stratagene Bgl II/Bam H1 (SEQ ID NO: 106) pSPEXCAL2 6.4486 bp SPEX pCAL-C Stratagene Bam H1/Bam extracellular H1 domain (SEQ IDNO: 110) pSPEXCAL3 6.2 464 bp SPEX pCAL-C Stratagene Not I, Bgl II/fragment Hind III (SEQ ID NO: 111) pSPEXdeltaN 4.5 609 bp SPEX gene pCR2.1 Invitrogen BamHI, EcoRI/ (internal domain) TOPO BamHI, Eco RI (SEQID NO: 108) pSPEXIgTOPO 4.45 538 bp SPEX pCR 2.1 Invitrogen Bgl II/BamH1 external domain TOPO (SEQ ID NO: 112) p1017SPEX 9.3 1276 bp SPEX genep1017 Int. Immunol. Bgl II/Bam H1 (SEQ ID NO: 109) (1990) 2: 173-180SPEXYFP- 6.5 1722 bp SPEX SEQ pMIGR1- Blood (1998) Not I, Bgl II/ MIGR1NOT (SEQ ID NO: 107) NOT Linker 92: 3780 Hind III pSPEX-Ig 8 538 bp SPEXpL-Selectin- Kpn I/Bam HI external domain Fc (SEQ ID NO: 112) pSPEXIg.B4.5 537 bp mSPEXb pCR 2.1 Invitrogen Bgl II/Bam H1 TOPO external domainTOPO (SEQ ID NO: 113)

[0171] Each of the vectors in Table 6 above are produced using standardcloning methods known to one of ordinary skill in the art. “BackboneDesignation” refers to the “vector sequence”. TABLE 7 PREFERRED USE OFCERTAIN SPEX VECTORS SPEX Vector Designation Preferred Use pSPEX-YFPExpression of SPEX-YFP fusion. YFP is the yellow fluorescent proteindetection marker. SPEXYFP-BABEPURO Retroviral vector for SPEXexpression. pSPEX-Bgl II Cloning of SPEX polynucleotides. pSPEX-BglII-BABEpuro Retroviral vector for SPEX expression. pSPEXCAL Bacterialexpression of SPEX polypeptides. pSPEXCAL2 Bacterial expression of SPEXpolypeptides. pSPEXCAL3 Bacterial expression of SPEX polypeptides.pSPEXdeltaN Vector encoding the intracellular domain of SPEX.pSPEXIgTOPO Cloning and Expression of a SPEX External Domain-IgG1chimera. p1017SPEX Expression of SPEX polypeptides in thymocytes withLck, a thymocyte specific promoter. SPEXYFP-MIGR1 NOT Retroviral vectorfor SPEX expression. Expression of SPEX in lymphocytes. pSPEX-IgExpression of a SPEX external domain-IgG1 fusion. Provides a SPEXexternal domain with enhanced solubility. pSPEXIg.B TOPO Cloning of SPEXexternal domain to make an in frame fusion with IgG1.

[0172] 4. Antibodies

[0173] One embodiment of the present invention provides an antibody thatimmunoreacts with a SPEX polypeptide. The term “antibody” is meant toinclude any form of antibody, including intact antibodies moleculesand/or an immunologically active portion of an antibody molecule.Antibodies and active fragments thereof are well known in the art, forexample: IgG, IgM, IgE, polyclonal, monoclonal, Fab, Fab′, F(ab′)₂,F(v), single chain antibody (SCA), single chain Fab, humanized, hybrid,and the like). Antibodies provided in the present invention immunoreactwith one or more portions of a SPEX polypeptide. It is preferred thatthe antibody is isolated. It is still more preferred that the antibodycomprises a recombinant, chimeric, or otherwise non-naturally antibodyoccurring.

[0174] Anti-SPEX antibodies are useful, for example, in modulating SPEXsignal transduction activity, lymphocyte activation or metabolism, andthe immune response. Anti-SPEX antibodies are also useful as a detectionagent in diagnostics and kits useful in detecting, purifying, and/orsorting SPEX expressing lymphocyte populations.

[0175] A preferred antibody is an anti-SPEX monoclonal antibody,preferably a human or humanized anti-SPEX monoclonal antibody. Otherpreferred anti-SPEX antibodies specifically immunoreact with one of ahSPEX polypeptide, a mSPEX polypeptide, or the mSPEXb polypeptide (theallele of murine SPEX disclosed above). An anti-SPEX antibody thatimmunoreacts with SEQ ID NO:7, SEQ ID NO:9, and/or SEQ ID NO: 11 iscontemplated to immunoreact with all murine and human SPEX polypeptidesincluding a SPEX tyrosine based domain (alternately with mammalian SPEXpolypeptides which have a SPEX tyrosine based domain or an intracellulardomain. Other exemplary anti-SPEX antibodies immunoreact with a SPEXpolypeptide set forth in TABLE 1 and TABLE 2, TABLE 3, and TABLE 4. Incertain embodiments, the anti-SPEX antibody immunoreacts with a firstSPEX polypeptide, but does not immunoreact with a second SPEXpolypeptide. For example, an anti-SPEX antibody immunoreacts with a SPEXpolypeptide set forth in column 1 and/or column 2 of TABLE 1, but doesnot immunoreact with a SPEX polypeptide set forth in column 3 ofTABLE 1. In another example, an anti-SPEX antibody immunoreacts with aSPEX polypeptide set forth in column 1 of TABLE 2, but does notimmunoreact with a second SPEX polypeptide set forth in column 2 ofTABLE 2.

[0176] The antigenic immunoreactivities of certain preferred anti-SPEXantibodies along with exemplary sequences, set forth by sequenceidentifier, are provided in TABLE 8, below. TABLE 8 PREFERRED ANTI-SPEXANTIBODIES IMMUNOREACT WITH THE FOLLOWING ANTIGENS Exemplary hSPEXExemplary mSPEX Polypeptide Polypeptide Optionally not Antigen(s)Antigen(s) Immunoreactive with SPEX Antigen (SEQ ID NO) (SEQ ID NO) (SEQID NO) Extracellular Domain 12, 13 54, 55, 85, 86 16, 17, 18, 58, 59, 60Extracellular/Transmembrane 14, 15 56, 57 16, 17, 58, 59 Ig Like Domain3 45, 88 16, 17, 18, 58, 59, 60 SPEX 20, 21 62, 63 Exemplary SPEX Fusion98 99, 100 16, 17, 18, 58, 59, 60 Polypeptide Intracellular Domain 7, 9,11, 16, 17, 18 7, 9, 11, 58, 59, 60 3, 45, 12, 13, 54, 55Intracellular/Transmembrane 18 60 3, 45, 12, 13, 54, 55, 14, 15, 56, 57

[0177] One embodiment provides an antibody, preferably monoclonal, thatimmunoreacts with an extracellular domain of a SPEX polypeptide and iscapable of modulating the proliferation or other metabolism oflymphocytes. More preferably, the antibody comprises a humanizedmonoclonal antibody. In one embodiment, administration of an antibodythat immunoreacts with a SPEX extracellular domain, preferably the Iglike domain, to lymphocytes including an expressed SPEX polypeptide;inhibits proliferation and/or differentiation of the lymphocyte.

[0178] Optionally an anti-SPEX antibody may include a binding domain fora second epitope, for instance a different SPEX epitope or even anon-SPEX epitope (e.g., a multivalent antibody capable of binding one ormore different epitopes).

[0179] In one embodiment, an anti-SPEX antibody immunoreacts with ahuman SPEX polypeptide, but does not substantially immunoreact with amouse SPEX polypeptide. Optionally, an anti-SPEX antibody immunoreactswith a mouse SPEX polypeptide, but does not substantially immunoreactwith a human SPEX polypeptide. Such antibodies are useful, for example,to distinguish human and mouse SPEX polypeptide sequences.

[0180] Methods of manufacturing an antibody given a specific antigen arewell known in the art. The present invention provides SPEX antigens(i.e., immunogen) useful for manufacture of anti-SPEX antibodies. Thus,in light of the present disclosure, one of ordinary skill in the art isable to manufacture an anti-SPEX antibody of the present invention.Antibodies are commonly manufactured, for example: in animals (e.g.,rabbit, mouse, hamster, sheep, goat, horse, bovine); in cells, primarilycell culture, (e.g., bacteria, plant, algae, insect, mammalian, murine,hybridoma, and human cells); by phage display; and by epitope cloninginto antibody scaffold vectors and gene transfer into any of a varietyof cell types (e.g., bacteria, plant, algae, insect, mammalian, murine,and human). Accordingly, the present invention also provides a cellcapable of producing an anti-SPEX antibody.

[0181] In certain embodiments, the anti-SPEX antibody includes adetectable tag. Examples of a detectable tag include a radioisotope(e.g., ¹²⁵I), a fluorescent molecule (e.g., SMCC-activatedBSA-Fluorescein, Prozyme, San Leandro, Calif.), biotinylation, myc tag,FLAG tag, and enzymes (e.g., peroxidase). Detectable tags and thelabeling or conjugation of an antibody with a desirable detectable tagis well known in the art. Typically, the detectable tag is combined withthe antibody by a covalent bond. Numerous commercial products areavailable for detectably labeling an antibody. It is preferred that thelabel does not interfere with structure and/or function of the variableportion of the antibody. Labeling in alternative areas of an antibodymolecule or active fragment thereof are well known in the art.

[0182] In certain embodiments, it is desirable that an anti-SPEXantibody immunoreacts with a first SPEX polypeptide, but does notimmunoreact with a second SPEX polypeptide. Monoclonal and polyclonalanti-SPEX antibodies having the present property can be prepared, forexample, using epitope specific SPEX antigens in the manufacture of theanti-SPEX antibody. If desired, an antibody population can be screenedto subtract or otherwise remove molecules in the population having anundesirable immunoreactivity (i.e., by subtraction screening on a matrixcontaining the undesirable antigen).

[0183] 5. Method of Screening for SPEX Binding Partners

[0184] One embodiment provides a method of screening candidate moleculesto identify one or more SPEX binding partners, comprising: 1) contactinga candidate molecule with a SPEX polypeptide; 2) determining whether ornot the candidate molecule and the SPEX polypeptide form a bindingcomplex, wherein the formation of a binding complex indicates that thecandidate molecule is a SPEX binding partner; and 3) repeating steps 1)and 2) until the SPEX binding partner is identified. Preferred candidatebinding partner molecules include small molecule organic compounds andpolypeptides. Preferred SPEX polypeptide binding targets include a SPEXtyrosine based domain (e.g., SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:11)or a SPEX Ig like domain (e.g., SEQ ID NO:3, SEQ ID NO:45, or SEQ IDNO:88).

[0185] In light of the present invention and the knowledge in the art,the determination of whether or not the candidate molecule and the SPEXpolypeptide form a binding complex can be made using a variety ofbinding assays well known in the art, so the invention is not limited bythe particular binding assay. For example, the binding complex can bedetected by a molecular weight increase compared to the SPEX polypeptidealone with capture of the binding complex using an anti-SPEX antibody.The SPEX polypeptide may be engineered to include an affinity tag or adetectable label such that the binding complex can be identified througha change in molecular weight or migration through a gel, or directdetection of the SPEX polypeptide in a complex. Certain candidatemolecules may have a known detectable property, such as an antibodyimmunoreactive site (epitope), fluorescent property, or affinity tag(natural or engineered) such that the candidate molecule can be directlydetected in a binding complex with a SPEX polypeptide.

[0186] 6. Method of Isolating a SPEX Polypeptide

[0187] One embodiment provides a method of purifying a SPEX polypeptidefrom a biological sample containing the SPEX polypeptide, comprising:contacting the biological sample with an affinity matrix having ananti-SPEX antibody attached to the matrix to produce an immunocomplexincluding the SPEX polypeptide and the antibody attached to the matrix;separating the remainder of the biological sample from the matrix;separating the SPEX polypeptide from the anti-SPEX antibody; andcollecting the SPEX polypeptide, thereby obtaining the purified SPEXpolypeptide.

[0188] 7. Method of Regulating an Activity of Lymphocytes

[0189] One embodiment provides a method of modulating a metabolism(preferably proliferation or activation) of a lymphocyte that expressesa SPEX receptor, comprising contacting the lymphocyte with an anti-SPEXantibody (preferably a monoclonal antibody) that immunoreacts with anextracellular domain of the SPEX polypeptide. Certain embodimentsdisclose that SPEX receptor activity modulates, preferably inhibitsmetabolism, and more preferably inhibits proliferation, of lymphocytesthat express the SPEX receptor. An antibody that immunoreacts with theextracellular domain of the SPEX polypeptide, preferably the Ig likedomain, is disclosed in certain embodiments herein to inhibit SPEXactivity providing a release from SPEX induced suppression of themetabolism thus the antibody provides a corresponding increase in themetabolism of the SPEX expressing lymphocyte. Useful antibodies includethe rat anti-SPEX antibodies PK3, PK18, and PK23 (or immunoreactivefragments thereof). Preferred antibodies include humanized or humanantibodies that immunoreact with an extracellular domain of a SPEXpolypeptide. Preferred lymphocytes include B cells and T cells. Incertain preferred embodiments, the antibody immunoreacts with an aminoacid sequence set forth in SEQ ID NO:13, SEQ ID NO:55, or SEQ ID NO:85.The lymphocytes can be in an in vitro culture or in vivo. Lymphocytesare contacted with the antibody in vivo, for example, by administeringthe antibody or immunoreactive fragment thereof (e.g., by injection) toa human, or non-human mammal.

EXAMPLES 1. Identify and Clone mSPEX cDNA

[0190] Positive selection is a developmental process in which immaturethymocytes receive maturation signals as a consequence of T cell antigenreceptor (TCR) recognition of MHC/self-peptide complexes expressed bythymic stroma. Exposure of immature thymocytes to low concentrations ofthe pharmacologic activators phorbol ester (e.g., PMA) and/or ionomycininduces the survival and differentiation of double positive (DP)thymocytes in vitro and provides an accepted model system of positiveselection of thymocytes in vivo.

[0191] Using nucleic acid microarrays, the inventors identified changesin nucleic acid expression during positive selection of thymocytesrelative to unstimulated thymocytes. TCRα-chain deficient thymocytes(murine) were cultured in medium in the presence or absence of 0.2 ng/mlPMA and 0.2 mg/ml ionomycin providing activated and non-activatedthymocytes respectively. TCRα-chain deficient thymocytes are blocked atthe DP stage of development due to a genetic mutation. Gene expressionin these cells, after stimulation, is characteristic of developingthymocytes. After incubating the cells for 6 hours, the poly-A+ RNA wasisolated using RNeasy RNA and Oligotex mRNA kits (Qiagen). The poly-A+RNA was subjected to comparative cDNA array analysis using the Mouse1.02 Array per manufacturer's instructions (Incyte Genomics). Signalanalysis of the microarrays was performed using GEMTools analysissoftware (Incyte Genomics).

[0192] The inventors selected two sequences for further study based uponthe ratio of expression of the sequences in the stimulated thymocytesover the unstimulated thymocytes. The ratio of expression was determinedfrom the normalized signals of hybridization of each labeled cDNA to theESTs AA184189 and AA177302, respectively, which ESTs were included onthe Mouse Array. The expression of each sequence in the stimulatedthymocytes was increased 9.1 fold and 6.6 fold for sequences hybridizingto EST AA184189 and EST AA177302, respectively. These ESTs on the Mouse1.02 Array originated from a murine library called the Soares mouse3NbMS library which in turn was derived from spleens of 4 week oldC57BL/6J mice.

[0193] The inventors determined that the ESTs AA184189 and AA177302 arelinked using BLAST software (NCBI) to compare the sequences of the ESTswith a database of murine nucleic acid sequences (each EST correspondsto a common sequence in the murine sequence database). Thus, ESTsAA184189 and AA177302 are determined to be part of a more completenucleic acid. Using the 5′-most EST (AA177302), the inventors cloned afull-length gene by rapid amplification of cDNA ends (SMART RACE cDNAamplification kit, Clontech) using cDNA prepared from stimulatedthymocytes as a template in the reaction.

[0194] The present gene is designated herein as the mouse spleenexpressed gene (mSPEX gene). An exemplary mSPEX nucleic acid is setforth in SEQ ID NO:105 and includes a coding region set forth in SEQ IDNO:84. A BLAST search of GenBank reveals that the SPEX cDNA encodes anovel protein. An exemplary mSPEX polypeptide encoded by an mSPEXnucleic acid is set forth in SEQ ID NO:63. A signal sequence (e.g., SEQID NO:43) is typically cleaved during cellular processing to form anexemplary mature mSPEX polypeptide set forth in SEQ ID NO:62. Usingsequence comparison between the mSPEX polypeptide and the Pfam family ofproteins database (NCBI), the inventors identified an immunoglobulinlike domain in a predicted extracellular portion of the polypeptide;however, the inventors observed that SPEX lacks close homology to anyparticular immunoglobulin containing superfamily member.

2. Identify and Clone hSPEX cDNA

[0195] Using sequence comparisons of the mSPEX sequence to humansequences in the GenBank database the inventors determine that the humanESTs AI792952 and AA931122 align with the mSPEX polynucleotide and thatthe human ESTs correspond to a single human clone in the GenBankdatabase. A bacterial stab (AI792952) containing the human clone, aswell as numerous contaminating clones, was purchased from the IMAGEConsortium (Lawrence Livermore National Laboratory, Livermore, Calif.).A human gene sequence homologous to mSPEX is purified from the stab,subcloned, and sequenced. An exemplary hSPEX nucleic acid sequence isset forth in SEQ ID NO: 104 and includes a coding region set forth inSEQ ID NO:42. An exemplary hSPEX polypeptide encoded by an hSPEX nucleicacid sequence is set forth in SEQ ID NO:21. A signal sequence (e.g., SEQID NO: 1) is typically cleaved during cellular processing of hSPEXprotein to form an exemplary mature hSPEX polypeptide set forth in SEQID NO:20.

3. Anti-mSPEX Monoclonal Antibody

[0196] An isolated expression vector construct (referred to herein asp-mSPEX-Ig) comprising a coding region encoding a fusion protein, thefusion having a mSPEX extracellular domain (including the SPEX signalsequence) and the hinge, CH2, and CH3 domains of human IgG1 is prepared.The p-mSPEX-Ig vector is transfected into 293 cells in culture. Thep-mSPEX-Ig transfected 293 cells express and secrete the mSPEX-Ig fusionprotein as assessed by a Western blot probed with ani-humanIgG1antibody. The mSPEX-Ig fusion protein is purified by protein A affinitychromatography from supernatants of transfected 293 cells.

[0197] Rats are immunized in the base of the tail with the purifiedrecombinant mSPEX-Ig emulsified in CFA to produce monoclonal antibodies.Procedures and techniques for the production of antibodies, includingmonoclonal antibodies, to a given antigen are well known in the art. Themedial iliac lymph nodes are harvested two weeks later and fused withYB2/0 cells by standard methods. Antibodies thus produced are screenedfor specific immunoreactivity with mSPEX by FACS using DPK and 293 cellsthat are transfected to express a cell surface mSPEX-YFP fusion protein(YFP is an abbreviation for yellow fluorescent protein). Screeningcontinues until one or more anti-mSPEX monoclonal antibody is identifiedin the screening process. The YFP tag allows verification thattransfected cells express the mSPEX-YFP fusion protein at the cellsurface and allows the correlation of the relative level of cellularantibody binding to mSPEX-YFP fusion protein expression. Specifically,the DPK and 293 cells used for the screening procedure are transfectedwith an isolated expression vector construct made from cloning a mSPEXgene insert into the pEYEP-N1 vector (Clontech). The pEYEP-N1 vectorsupplies an YFP tag and expression of a mSPEX insert from pEYEP-N1results in the expression of a fusion protein including a mSPEXpolypeptide operably linked with a YFP tag. Three hybridomas PK3, PK18,and PK23 are obtained from this screen. The hybridomas are optionallyre-cloned and antibody is purified using standard methods.

[0198] The monoclonal antibody thus produced specifically immunoreactswith the extracellular domain of mSPEX. Three monoclonal antibodies thatimmunoreact with the extracellular domain of mSPEX are: PK3, PK18, andPK23; as referred to herein.

4. Anti-hSPEX Monoclonal Antibody

[0199] A monoclonal antibody that specifically reacts with theextracellular domain of a hSPEX polypeptide is prepared using themethods disclosed in Example 3, except that 1) a p-hSPEX-Ig expressionvector comprising a coding region encoding a fusion protein, the fusionhaving a mSPEX extracellular domain (including the SPEX signal sequence)and the hinge, CH2, and CH3 domains of human IgG1 is prepared and usedto produce monoclonal antibodies in rats and 2) a hSPEX polynucleotideencoding the extracellular domain of a hSPEX polypeptide is cloned intoa pEYEP-N1 vector and expressed in DPK and 293 cells to screen formonoclonal antibodies using the FACS-YFP procedure disclosed above. Themonoclonal antibody produced specifically immunoreacts with theextracellular domain of hSPEX.

5. Map Kinase Signaling Stimulates SPEX Expression

[0200] The expression of mSPEX and hSPEX mRNAs is compared in culturedDP thymocytes in the presence or absence of stimulation with PMA andionomycin and in the presence or absence of a MEK inhibitor (10 μMU0126). The increase of SPEX mRNA expression (in both murine and humancells) observed with PMA and ionomycin stimulation is inhibited by theMEK inhibitor by approximately 70% in one experiment. These data areconsistent with MAP kinase signaling leading to stimulation of SPEX mRNAexpression.

6. Isolation of Cellular mSPEX

[0201] The mSPEX protein is isolated under reducing and non-reducingconditions from murine splenocyte whole cell lysates by affinitychromatography using a matrix bound anti-mSPEX monoclonal antibody. Thematrix bound antibody is contacted with the cell lysates underconditions sufficient for binding of mSPEX polypeptide in the lysateswith the antibody thereby forming a reaction complex including thematrix, the anti-hSPEX antibody, and hSPEX polypeptide (immunoreactionbuffers and other conditions, such as incubation time and temperature,are well known in the art and are applicable here). The matrix is washedto remove non-mSPEX contaminants. The mSPEX polypeptide is released fromthe matrix-antibody-mSPEX complex and the mSPEX polypeptide is collectedin purified form. The presence of purified mSPEX polypeptide isconfirmed by electrophoresis on SDS-PAGE gels using silver staining todetect biological contents of the samples. The mSPEX polypeptide isessentially the only material present in the samples, appearing as a 35kDa and 37 kDa doublet as detected by silver staining. The identity ofeach band of the doublet is confirmed to be mSPEX by epitope tagging oftransfected cells and by Western blotting using an anti-mSPEX monoclonalantibody.

7. Isolation of Cellular hSPEX

[0202] The hSPEX protein is isolated under reducing and non-reducingconditions from human splenocytes cell culture lysates by affinitychromatography using a matrix bound anti-hSPEX monoclonal antibody. Thematrix bound antibody is contacted with the cell lysates underconditions sufficient for binding of hSPEX polypeptide in the lysateswith the antibody thereby forming a reaction complex including thematrix, the anti-hSPEX antibody, and hSPEX polypeptide. The matrix iswashed to remove non-hSPEX contaminants. The hSPEX polypeptide isreleased from the matrix-antibody-hSPEX complex and the hSPEXpolypeptide is collected in purified form. The presence of purifiedhSPEX polypeptide in the sample is confirmed by electrophoresis onSDS-PAGE gels using silver staining to detect the contents of thesample. The hSPEX polypeptide is essentially the only material presentin the samples as detected by silver staining. The identity of the hSPEXpolypeptide is confirmed by Western blotting using an anti-hSPEXmonoclonal antibody.

8. Tissue Distribution of SPEX Protein

[0203] The tissue distribution of SPEX gene expression is examined inmurine brain, heart, kidney, liver, lung, muscle, skin, small intestine,spleen, stomach, testis, and thymus tissues by Northern blot analysis ofcellular mRNA. Essentially no SPEX mRNA is detected by Northern analysisin murine brain or testis. Low level expression of SPEX is observed inheart, kidney, muscle, and skin tissues. Intermediate level expressionof SPEX is observed in liver, lung, small intestine, and stomachtissues. High level expression of SPEX is observed in the thymus andspleen tissues with about 2 fold greater expression in the spleen tissuecompared to the thymus tissue.

[0204] The distribution of SPEX mRNA expression in lymphocytes isevaluated in murine B-lymphocytes, CD4+ thymocytes, and CD8+ thymocytes.Expression of SPEX in thymocytes is determined by 3 color staining thethymocytes for CD4, CD8, and SPEX and analyzing the expression patternsby flow cytometry. The SPEX mRNA expression is higher on CD4 T cellscompared to CD8 T cells. In one experiment the CD4 T cell SPEX mRNAexpression over CD8 T cell SPEX mRNA expression was approximately 2:1.The expression of SPEX protein is also higher on CD4 T cells compared tothe CD8 lineage, although both population of cells express the protein.These data are consistent with SPEX signaling participating in lineagecommitment of T cells.

[0205] Expression of SPEX mRNA in B-cells, CD4 T-cells, and CD8 T-cellsis evaluated by 3 color staining murine spleen cells and analyzing theexpression patterns by FACS. Again, SPEX is expressed on B and T cellswith an typical order of expression level from highest to lowest being Bcells, followed by CD4 T cells, and then by CD8 cells.

9. Regulation of T Cell Response With Anti-SPEX Antibody

[0206] This example detects the modulation of a T cell metabolism bycontacting a T cell with an anti-mSPEX antibody. CD4 T cells arepurified by magnetic bead depletion from lymph nodes of AND TCRtransgenic mice. These T cells have a well characterized antigenspecificity regarding CD4 and CD8 expression. In the present assay, theCD4 T cells are admixed with antigen presenting cells (APCs) and anantigen in the presence or absence of a SPEX specific monoclonalantibody. Cultures of each admixture are incubated for differentselected time periods and the T cell response is detected or measured. Achange in the T cell proliferation rate in the antibody treatedadmixtures compared to admixtures without the antibody is evidence ofmodulation of T cell metabolism resulting from the administration of theantibody which immunoreacts with an extracellular domain of SPEX.

[0207] The APCs in the above assay are prepared from irradiated spleencells of BALB.K mice according to standard techniques known in the art.The APCs have the appropriate MHC molecules to present antigen to theAND T cells, have the mSPEXb allele, and express the mSPEXb polypeptide;but do not have the SPEX allele or express a mSPEX polypeptide otherthan mSPEXb. The antibody used is PK18 which is an anti-mSPEX antibodywhich does not immunoreact with the mSPEXb polypeptide. The PK18antibody immunoreacts with the extracellular domain of the mSPEXpolypeptide. Thus, the APCs prepared from BALB.K mice do not immunoreactwith the PK18 antibody. T cells and APC are mixed in the presence ofvarious concentrations of antigen, in this example a peptide derivedfrom pigeon cytochrome c, in the presence or absence of PK18 anti-SPEXmonoclonal antibody, and cultured for various time periods.Proliferation is measured by ³H thymidine uptake at various time periodsafter the start of culture.

10. Inhibition of T Cell Activation With an Anti-SPEX Antibody

[0208] This example demonstrates a method of inhibiting T cellactivation by contacting a T cell containing a SPEX polypeptide with anantibody that immunoreacts with the SPEX polypeptide. LN T cells areseparately purified from B10.BR (which have a mSPEX polypeptide) andBALB.K mice (which have a mSPEXb polypeptide). The T cells are culturedin the presence of 3H-TdR (tritiated thymidine) in 96-well plates coatedwith equal amounts of anti-CD3 and rat Ig or equal amounts of anti-CD3and monoclonal antibody PK18. The anti-CD3 antibody is a knownstimulator of T cell activation. The PK18 antibody immunoreacts with themSPEX polypeptide of the B10.BR T cells, but does not immunoreact withthe mSPEXb polypeptide of the BALB.K T cells.

[0209] Results are shown in FIG. 3. 3H-TdR incorporation into T cellsreacted with anti-CD3 and rat Ig is depicted as open symbols in FIG. 3.3H-TdR incorporation into T cells reacted with anti-CD3 and PK18 isdepicted as closed symbols in FIG. 3. The data are expressed as the mean3H-TdR incorporation for triplicate cultures (+/− standard deviation(SD)) minus the cpm of 3H-TdR incorporation of cultures of T cells alone(which was less than 1500 cpm). The incorporation of 3H-TdR into the Tcells is correlated with both proliferation of the T cells and with Tcell activation.

[0210] Panels 1 and 2 (two different experiments) of FIG. 3 demonstratethat the activation of T cells from B10 mice is inhibited when the Tcells are contacted with the antibody PK18 which immunoreacts with themSPEX of the B10 mouse T cells. Panels 3 and 4 (two differentexperiments) of FIG. 3 demonstrate that the activation of T cells fromBALB.K mice is not significantly modulated when the T cells arecontacted with the antibody PK18 which does not immunoreact with themSPEXb of the BALB.K mouse T cells.

[0211] The PK18 antibody immunoreacts with the extracellular domain ofthe mSPEXb polypeptide. Thus, the data depicted in FIG. 3 demonstrate amethod of inhibiting T cell activation by contacting a T cell containinga SPEX polypeptide with an antibody that immunoreacts with the SPEXpolypeptide, preferably the extracellular domain of the SPEXpolypeptide, and more preferably the Ig like domain of the SPEXpolypeptide.

11. Inhibition of B Cell Activation With an Anti-SPEX Antibody

[0212] This example demonstrates a method of inhibiting B cellactivation by contacting a B cell containing a SPEX polypeptide with anantibody that immunoreacts with the SPEX polypeptide. Example 10 isrepeated except that B cells are purified from the B10.BR and BALB.Kmice. It is found that B cell activation is inhibited by contacting theB cells of the B10, but not the BALB mice with the PK18 antibody. Thus,the data demonstrate a method of inhibiting B cell activation bycontacting a B cell containing a SPEX polypeptide with an antibody thatimmunoreacts with the SPEX polypeptide, preferably the extracellulardomain of the SPEX polypeptide, and more preferably the Ig like domainof the SPEX polypeptide.

12. Murine Anti-MSPEX Monoclonal Antibody

[0213] BALB/c mice are immunized (50% g in CFA) and boosted (3 to 5times, 50% g in IFA) with recombinant soluble SPEX-Ig fusion protein(including the extracellular domain of SPEX as discussed above). Theimmunogen is derived from B6 mice (containing the mSPEX allele, not themSPEXb allele) and thus generates allele specific antibodies in BALB/cmice. Hybridomas are produced by standard means from splenocytes ofimmunized animals. Monoclonal antibodies are screened by differentialbinding to parental and transfected cells expressing SPEX-YFP fusionprotein. Three monoclonal antibodies, PJ16, PJ19 and PJ196, are selectedfor additional analysis. The isotype of each antibody is IgG1κ. ThePJ19, PJ16, and PJ196 antibodies have identical staining patterns to thePK series of rat monoclonal antibodies (see above), includingspecifically immunoreacting with mSPEX (but not mSPEXb) expressed by B6cells, but not BALB/c derived cells (which express mSPEXb, but notmSPEX).

[0214] Various modifications of the invention, in addition to theembodiments provided herein, will become apparent to those skilled inthe art from the foregoing description and fall within the scope of theclaims.

1 113 1 30 PRT Homo sapiens 1 Met Lys Thr Leu Pro Ala Met Leu Gly ThrGly Lys Leu Phe Trp Val 1 5 10 15 Phe Phe Leu Ile Pro Tyr Leu Asp IleTrp Asn Ile His Gly 20 25 30 2 20 PRT Homo sapiens 2 Lys Glu Ser Cys AspVal Gln Leu Tyr Ile Lys Arg Gln Ser Glu His 1 5 10 15 Ser Ile Leu Ala 203 65 PRT Homo sapiens 3 Gly Asp Pro Phe Glu Leu Glu Cys Pro Val Lys TyrCys Ala Asn Arg 1 5 10 15 Pro His Val Thr Trp Cys Lys Leu Asn Gly ThrThr Cys Val Lys Leu 20 25 30 Glu Asp Arg Gln Thr Ser Trp Lys Glu Glu LysAsn Ile Ser Phe Phe 35 40 45 Ile Leu His Phe Glu Pro Val Leu Pro Asn AspAsn Gly Ser Tyr Arg 50 55 60 Cys 65 4 42 PRT Homo sapiens 4 Ser Ala AsnPhe Gln Ser Asn Leu Ile Glu Ser His Ser Thr Thr Leu 1 5 10 15 Tyr ValThr Asp Val Lys Ser Ala Ser Glu Arg Pro Ser Lys Asp Glu 20 25 30 Met AlaSer Arg Pro Trp Leu Leu Tyr Ser 35 40 5 21 PRT Homo sapiens 5 Leu LeuPro Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys 1 5 10 15 LeuPhe Cys Cys Leu 20 6 44 PRT Homo sapiens 6 Arg Arg His Gln Gly Lys GlnAsn Glu Leu Ser Asp Thr Ala Gly Arg 1 5 10 15 Glu Ile Asn Leu Val AspAla His Leu Lys Ser Glu Gln Thr Glu Ala 20 25 30 Ser Thr Arg Gln Asn SerGln Val Leu Leu Ser Glu 35 40 7 8 PRT Homo sapiens 7 Thr Gly Ile Tyr AspAsn Asp Pro 1 5 8 23 PRT Homo sapiens 8 Asp Leu Cys Phe Arg Met Gln GluGly Ser Glu Val Tyr Ser Asn Pro 1 5 10 15 Cys Leu Glu Glu Asn Lys Pro 209 9 PRT Homo sapiens 9 Gly Ile Val Tyr Ala Ser Leu Asn His 1 5 10 14 PRTHomo sapiens 10 Ser Val Ile Gly Leu Asn Ser Arg Leu Ala Arg Asn Val Lys1 5 10 11 13 PRT Homo sapiens 11 Glu Ala Pro Thr Glu Tyr Ala Ser Ile CysVal Arg Ser 1 5 10 12 127 PRT Homo sapiens 12 Lys Glu Ser Cys Asp ValGln Leu Tyr Ile Lys Arg Gln Ser Glu His 1 5 10 15 Ser Ile Leu Ala GlyAsp Pro Phe Glu Leu Glu Cys Pro Val Lys Tyr 20 25 30 Cys Ala Asn Arg ProHis Val Thr Trp Cys Lys Leu Asn Gly Thr Thr 35 40 45 Cys Val Lys Leu GluAsp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn 50 55 60 Ile Ser Phe Phe IleLeu His Phe Glu Pro Val Leu Pro Asn Asp Asn 65 70 75 80 Gly Ser Tyr ArgCys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser 85 90 95 His Ser Thr ThrLeu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg 100 105 110 Pro Ser LysAsp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Ser 115 120 125 13 157 PRTHomo sapiens 13 Met Lys Thr Leu Pro Ala Met Leu Gly Thr Gly Lys Leu PheTrp Val 1 5 10 15 Phe Phe Leu Ile Pro Tyr Leu Asp Ile Trp Asn Ile HisGly Lys Glu 20 25 30 Ser Cys Asp Val Gln Leu Tyr Ile Lys Arg Gln Ser GluHis Ser Ile 35 40 45 Leu Ala Gly Asp Pro Phe Glu Leu Glu Cys Pro Val LysTyr Cys Ala 50 55 60 Asn Arg Pro His Val Thr Trp Cys Lys Leu Asn Gly ThrThr Cys Val 65 70 75 80 Lys Leu Glu Asp Arg Gln Thr Ser Trp Lys Glu GluLys Asn Ile Ser 85 90 95 Phe Phe Ile Leu His Phe Glu Pro Val Leu Pro AsnAsp Asn Gly Ser 100 105 110 Tyr Arg Cys Ser Ala Asn Phe Gln Ser Asn LeuIle Glu Ser His Ser 115 120 125 Thr Thr Leu Tyr Val Thr Asp Val Lys SerAla Ser Glu Arg Pro Ser 130 135 140 Lys Asp Glu Met Ala Ser Arg Pro TrpLeu Leu Tyr Ser 145 150 155 14 148 PRT Homo sapiens 14 Lys Glu Ser CysAsp Val Gln Leu Tyr Ile Lys Arg Gln Ser Glu His 1 5 10 15 Ser Ile LeuAla Gly Asp Pro Phe Glu Leu Glu Cys Pro Val Lys Tyr 20 25 30 Cys Ala AsnArg Pro His Val Thr Trp Cys Lys Leu Asn Gly Thr Thr 35 40 45 Cys Val LysLeu Glu Asp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn 50 55 60 Ile Ser PhePhe Ile Leu His Phe Glu Pro Val Leu Pro Asn Asp Asn 65 70 75 80 Gly SerTyr Arg Cys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser 85 90 95 His SerThr Thr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg 100 105 110 ProSer Lys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Ser Leu 115 120 125Leu Pro Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys Leu 130 135140 Phe Cys Cys Leu 145 15 178 PRT Homo sapiens 15 Met Lys Thr Leu ProAla Met Leu Gly Thr Gly Lys Leu Phe Trp Val 1 5 10 15 Phe Phe Leu IlePro Tyr Leu Asp Ile Trp Asn Ile His Gly Lys Glu 20 25 30 Ser Cys Asp ValGln Leu Tyr Ile Lys Arg Gln Ser Glu His Ser Ile 35 40 45 Leu Ala Gly AspPro Phe Glu Leu Glu Cys Pro Val Lys Tyr Cys Ala 50 55 60 Asn Arg Pro HisVal Thr Trp Cys Lys Leu Asn Gly Thr Thr Cys Val 65 70 75 80 Lys Leu GluAsp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn Ile Ser 85 90 95 Phe Phe IleLeu His Phe Glu Pro Val Leu Pro Asn Asp Asn Gly Ser 100 105 110 Tyr ArgCys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser His Ser 115 120 125 ThrThr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg Pro Ser 130 135 140Lys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Ser Leu Leu Pro 145 150155 160 Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys Leu Phe Cys165 170 175 Cys Leu 16 67 PRT Homo sapiens 16 Thr Gly Ile Tyr Asp AsnAsp Pro Asp Leu Cys Phe Arg Met Gln Glu 1 5 10 15 Gly Ser Glu Val TyrSer Asn Pro Cys Leu Glu Glu Asn Lys Pro Gly 20 25 30 Ile Val Tyr Ala SerLeu Asn His Ser Val Ile Gly Leu Asn Ser Arg 35 40 45 Leu Ala Arg Asn ValLys Glu Ala Pro Thr Glu Tyr Ala Ser Ile Cys 50 55 60 Val Arg Ser 65 17111 PRT Homo sapiens 17 Arg Arg His Gln Gly Lys Gln Asn Glu Leu Ser AspThr Ala Gly Arg 1 5 10 15 Glu Ile Asn Leu Val Asp Ala His Leu Lys SerGlu Gln Thr Glu Ala 20 25 30 Ser Thr Arg Gln Asn Ser Gln Val Leu Leu SerGlu Thr Gly Ile Tyr 35 40 45 Asp Asn Asp Pro Asp Leu Cys Phe Arg Met GlnGlu Gly Ser Glu Val 50 55 60 Tyr Ser Asn Pro Cys Leu Glu Glu Asn Lys ProGly Ile Val Tyr Ala 65 70 75 80 Ser Leu Asn His Ser Val Ile Gly Leu AsnSer Arg Leu Ala Arg Asn 85 90 95 Val Lys Glu Ala Pro Thr Glu Tyr Ala SerIle Cys Val Arg Ser 100 105 110 18 132 PRT Homo sapiens 18 Leu Leu ProLeu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys 1 5 10 15 Leu PheCys Cys Leu Arg Arg His Gln Gly Lys Gln Asn Glu Leu Ser 20 25 30 Asp ThrAla Gly Arg Glu Ile Asn Leu Val Asp Ala His Leu Lys Ser 35 40 45 Glu GlnThr Glu Ala Ser Thr Arg Gln Asn Ser Gln Val Leu Leu Ser 50 55 60 Glu ThrGly Ile Tyr Asp Asn Asp Pro Asp Leu Cys Phe Arg Met Gln 65 70 75 80 GluGly Ser Glu Val Tyr Ser Asn Pro Cys Leu Glu Glu Asn Lys Pro 85 90 95 GlyIle Val Tyr Ala Ser Leu Asn His Ser Val Ile Gly Leu Asn Ser 100 105 110Arg Leu Ala Arg Asn Val Lys Glu Ala Pro Thr Glu Tyr Ala Ser Ile 115 120125 Cys Val Arg Ser 130 19 239 PRT Homo sapiens 19 Gly Asp Pro Phe GluLeu Glu Cys Pro Val Lys Tyr Cys Ala Asn Arg 1 5 10 15 Pro His Val ThrTrp Cys Lys Leu Asn Gly Thr Thr Cys Val Lys Leu 20 25 30 Glu Asp Arg GlnThr Ser Trp Lys Glu Glu Lys Asn Ile Ser Phe Phe 35 40 45 Ile Leu His PheGlu Pro Val Leu Pro Asn Asp Asn Gly Ser Tyr Arg 50 55 60 Cys Ser Ala AsnPhe Gln Ser Asn Leu Ile Glu Ser His Ser Thr Thr 65 70 75 80 Leu Tyr ValThr Asp Val Lys Ser Ala Ser Glu Arg Pro Ser Lys Asp 85 90 95 Glu Met AlaSer Arg Pro Trp Leu Leu Tyr Ser Leu Leu Pro Leu Gly 100 105 110 Gly LeuPro Leu Leu Ile Thr Thr Cys Phe Cys Leu Phe Cys Cys Leu 115 120 125 ArgArg His Gln Gly Lys Gln Asn Glu Leu Ser Asp Thr Ala Gly Arg 130 135 140Glu Ile Asn Leu Val Asp Ala His Leu Lys Ser Glu Gln Thr Glu Ala 145 150155 160 Ser Thr Arg Gln Asn Ser Gln Val Leu Leu Ser Glu Thr Gly Ile Tyr165 170 175 Asp Asn Asp Pro Asp Leu Cys Phe Arg Met Gln Glu Gly Ser GluVal 180 185 190 Tyr Ser Asn Pro Cys Leu Glu Glu Asn Lys Pro Gly Ile ValTyr Ala 195 200 205 Ser Leu Asn His Ser Val Ile Gly Leu Asn Ser Arg LeuAla Arg Asn 210 215 220 Val Lys Glu Ala Pro Thr Glu Tyr Ala Ser Ile CysVal Arg Ser 225 230 235 20 259 PRT Homo sapiens 20 Lys Glu Ser Cys AspVal Gln Leu Tyr Ile Lys Arg Gln Ser Glu His 1 5 10 15 Ser Ile Leu AlaGly Asp Pro Phe Glu Leu Glu Cys Pro Val Lys Tyr 20 25 30 Cys Ala Asn ArgPro His Val Thr Trp Cys Lys Leu Asn Gly Thr Thr 35 40 45 Cys Val Lys LeuGlu Asp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn 50 55 60 Ile Ser Phe PheIle Leu His Phe Glu Pro Val Leu Pro Asn Asp Asn 65 70 75 80 Gly Ser TyrArg Cys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser 85 90 95 His Ser ThrThr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu Arg 100 105 110 Pro SerLys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr Ser Leu 115 120 125 LeuPro Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr Cys Phe Cys Leu 130 135 140Phe Cys Cys Leu Arg Arg His Gln Gly Lys Gln Asn Glu Leu Ser Asp 145 150155 160 Thr Ala Gly Arg Glu Ile Asn Leu Val Asp Ala His Leu Lys Ser Glu165 170 175 Gln Thr Glu Ala Ser Thr Arg Gln Asn Ser Gln Val Leu Leu SerGlu 180 185 190 Thr Gly Ile Tyr Asp Asn Asp Pro Asp Leu Cys Phe Arg MetGln Glu 195 200 205 Gly Ser Glu Val Tyr Ser Asn Pro Cys Leu Glu Glu AsnLys Pro Gly 210 215 220 Ile Val Tyr Ala Ser Leu Asn His Ser Val Ile GlyLeu Asn Ser Arg 225 230 235 240 Leu Ala Arg Asn Val Lys Glu Ala Pro ThrGlu Tyr Ala Ser Ile Cys 245 250 255 Val Arg Ser 21 289 PRT Homo sapiens21 Met Lys Thr Leu Pro Ala Met Leu Gly Thr Gly Lys Leu Phe Trp Val 1 510 15 Phe Phe Leu Ile Pro Tyr Leu Asp Ile Trp Asn Ile His Gly Lys Glu 2025 30 Ser Cys Asp Val Gln Leu Tyr Ile Lys Arg Gln Ser Glu His Ser Ile 3540 45 Leu Ala Gly Asp Pro Phe Glu Leu Glu Cys Pro Val Lys Tyr Cys Ala 5055 60 Asn Arg Pro His Val Thr Trp Cys Lys Leu Asn Gly Thr Thr Cys Val 6570 75 80 Lys Leu Glu Asp Arg Gln Thr Ser Trp Lys Glu Glu Lys Asn Ile Ser85 90 95 Phe Phe Ile Leu His Phe Glu Pro Val Leu Pro Asn Asp Asn Gly Ser100 105 110 Tyr Arg Cys Ser Ala Asn Phe Gln Ser Asn Leu Ile Glu Ser HisSer 115 120 125 Thr Thr Leu Tyr Val Thr Asp Val Lys Ser Ala Ser Glu ArgPro Ser 130 135 140 Lys Asp Glu Met Ala Ser Arg Pro Trp Leu Leu Tyr SerLeu Leu Pro 145 150 155 160 Leu Gly Gly Leu Pro Leu Leu Ile Thr Thr CysPhe Cys Leu Phe Cys 165 170 175 Cys Leu Arg Arg His Gln Gly Lys Gln AsnGlu Leu Ser Asp Thr Ala 180 185 190 Gly Arg Glu Ile Asn Leu Val Asp AlaHis Leu Lys Ser Glu Gln Thr 195 200 205 Glu Ala Ser Thr Arg Gln Asn SerGln Val Leu Leu Ser Glu Thr Gly 210 215 220 Ile Tyr Asp Asn Asp Pro AspLeu Cys Phe Arg Met Gln Glu Gly Ser 225 230 235 240 Glu Val Tyr Ser AsnPro Cys Leu Glu Glu Asn Lys Pro Gly Ile Val 245 250 255 Tyr Ala Ser LeuAsn His Ser Val Ile Gly Leu Asn Ser Arg Leu Ala 260 265 270 Arg Asn ValLys Glu Ala Pro Thr Glu Tyr Ala Ser Ile Cys Val Arg 275 280 285 Ser 2290 DNA Homo sapiens 22 atgaagacat tgcctgccat gcttggaact gggaaattattttgggtctt cttcttaatc 60 ccatatctgg acatctggaa catccatggg 90 23 60 DNAHomo sapiens 23 aaagaatcat gtgatgtaca gctttatata aagagacaat ctgaacactccatcttagca 60 24 201 DNA Homo sapiens 24 ggagatccct ttgaactagaatgccctgtg aaatactgtg ctaacaggcc tcatgtgact 60 tggtgcaagc tcaatggaacaacatgtgta aaacttgaag atagacaaac aagttggaag 120 gaagagaaga acatttcatttttcattcta cattttgaac cagtgcttcc taatgacaat 180 gggtcatacc gctgttctgc a201 25 120 DNA Homo sapiens 25 aattttcagt ctaatctcat tgaaagccactcaacaactc tttatgtgac agatgtaaaa 60 agtgcctcag aacgaccctc caaggacgaaatggcaagca gaccctggct cctgtatagt 120 26 63 DNA Homo sapiens 26ttacttcctt tggggggatt gcctctactc atcactacct gtttctgcct gttctgctgc 60 ctg63 27 132 DNA Homo sapiens 27 agaaggcacc aaggaaagca aaatgaactctctgacacag caggaaggga aattaacctg 60 gttgatgctc accttaagag tgagcaaacagaagcaagca ccaggcaaaa ttcccaagta 120 ctgctatcag aa 132 28 24 DNA Homosapiens 28 actggaattt atgataatga ccct 24 29 69 DNA Homo sapiens 29gacctttgtt tcaggatgca ggaagggtct gaagtttatt ctaatccatg cctggaagaa 60aacaaacca 69 30 27 DNA Homo sapiens 30 ggcattgttt atgcttccct gaaccat 2731 42 DNA Homo sapiens 31 tctgtcattg gactgaactc aagactggca agaaatgtaa aa42 32 39 DNA Homo sapiens 32 gaagcaccaa cagaatatgc atccatatgt gtgaggagt39 33 381 DNA Homo sapiens 33 aaagaatcat gtgatgtaca gctttatataaagagacaat ctgaacactc catcttagca 60 ggagatccct ttgaactaga atgccctgtgaaatactgtg ctaacaggcc tcatgtgact 120 tggtgcaagc tcaatggaac aacatgtgtaaaacttgaag atagacaaac aagttggaag 180 gaagagaaga acatttcatt tttcattctacattttgaac cagtgcttcc taatgacaat 240 gggtcatacc gctgttctgc aaattttcagtctaatctca ttgaaagcca ctcaacaact 300 ctttatgtga cagatgtaaa aagtgcctcagaacgaccct ccaaggacga aatggcaagc 360 agaccctggc tcctgtatag t 381 34 471DNA Homo sapiens 34 atgaagacat tgcctgccat gcttggaact gggaaattattttgggtctt cttcttaatc 60 ccatatctgg acatctggaa catccatggg aaagaatcatgtgatgtaca gctttatata 120 aagagacaat ctgaacactc catcttagca ggagatccctttgaactaga atgccctgtg 180 aaatactgtg ctaacaggcc tcatgtgact tggtgcaagctcaatggaac aacatgtgta 240 aaacttgaag atagacaaac aagttggaag gaagagaagaacatttcatt tttcattcta 300 cattttgaac cagtgcttcc taatgacaat gggtcataccgctgttctgc aaattttcag 360 tctaatctca ttgaaagcca ctcaacaact ctttatgtgacagatgtaaa aagtgcctca 420 gaacgaccct ccaaggacga aatggcaagc agaccctggctcctgtatag t 471 35 444 DNA Homo sapiens 35 aaagaatcat gtgatgtacagctttatata aagagacaat ctgaacactc catcttagca 60 ggagatccct ttgaactagaatgccctgtg aaatactgtg ctaacaggcc tcatgtgact 120 tggtgcaagc tcaatggaacaacatgtgta aaacttgaag atagacaaac aagttggaag 180 gaagagaaga acatttcatttttcattcta cattttgaac cagtgcttcc taatgacaat 240 gggtcatacc gctgttctgcaaattttcag tctaatctca ttgaaagcca ctcaacaact 300 ctttatgtga cagatgtaaaaagtgcctca gaacgaccct ccaaggacga aatggcaagc 360 agaccctggc tcctgtatagtttacttcct ttggggggat tgcctctact catcactacc 420 tgtttctgcc tgttctgctgcctg 444 36 534 DNA Homo sapiens 36 atgaagacat tgcctgccat gcttggaactgggaaattat tttgggtctt cttcttaatc 60 ccatatctgg acatctggaa catccatgggaaagaatcat gtgatgtaca gctttatata 120 aagagacaat ctgaacactc catcttagcaggagatccct ttgaactaga atgccctgtg 180 aaatactgtg ctaacaggcc tcatgtgacttggtgcaagc tcaatggaac aacatgtgta 240 aaacttgaag atagacaaac aagttggaaggaagagaaga acatttcatt tttcattcta 300 cattttgaac cagtgcttcc taatgacaatgggtcatacc gctgttctgc aaattttcag 360 tctaatctca ttgaaagcca ctcaacaactctttatgtga cagatgtaaa aagtgcctca 420 gaacgaccct ccaaggacga aatggcaagcagaccctggc tcctgtatag tttacttcct 480 ttggggggat tgcctctact catcactacctgtttctgcc tgttctgctg cctg 534 37 201 DNA Homo sapiens 37 actggaatttatgataatga ccctgacctt tgtttcagga tgcaggaagg gtctgaagtt 60 tattctaatccatgcctgga agaaaacaaa ccaggcattg tttatgcttc cctgaaccat 120 tctgtcattggactgaactc aagactggca agaaatgtaa aagaagcacc aacagaatat 180 gcatccatatgtgtgaggag t 201 38 333 DNA Homo sapiens 38 agaaggcacc aaggaaagcaaaatgaactc tctgacacag caggaaggga aattaacctg 60 gttgatgctc accttaagagtgagcaaaca gaagcaagca ccaggcaaaa ttcccaagta 120 ctgctatcag aaactggaatttatgataat gaccctgacc tttgtttcag gatgcaggaa 180 gggtctgaag tttattctaatccatgcctg gaagaaaaca aaccaggcat tgtttatgct 240 tccctgaacc attctgtcattggactgaac tcaagactgg caagaaatgt aaaagaagca 300 ccaacagaat atgcatccatatgtgtgagg agt 333 39 396 DNA Homo sapiens 39 ttacttcctt tggggggattgcctctactc atcactacct gtttctgcct gttctgctgc 60 ctgagaaggc accaaggaaagcaaaatgaa ctctctgaca cagcaggaag ggaaattaac 120 ctggttgatg ctcaccttaagagtgagcaa acagaagcaa gcaccaggca aaattcccaa 180 gtactgctat cagaaactggaatttatgat aatgaccctg acctttgttt caggatgcag 240 gaagggtctg aagtttattctaatccatgc ctggaagaaa acaaaccagg cattgtttat 300 gcttccctga accattctgtcattggactg aactcaagac tggcaagaaa tgtaaaagaa 360 gcaccaacag aatatgcatccatatgtgtg aggagt 396 40 717 DNA Homo sapiens 40 ggagatccct ttgaactagaatgccctgtg aaatactgtg ctaacaggcc tcatgtgact 60 tggtgcaagc tcaatggaacaacatgtgta aaacttgaag atagacaaac aagttggaag 120 gaagagaaga acatttcatttttcattcta cattttgaac cagtgcttcc taatgacaat 180 gggtcatacc gctgttctgcaaattttcag tctaatctca ttgaaagcca ctcaacaact 240 ctttatgtga cagatgtaaaaagtgcctca gaacgaccct ccaaggacga aatggcaagc 300 agaccctggc tcctgtatagtttacttcct ttggggggat tgcctctact catcactacc 360 tgtttctgcc tgttctgctgcctgagaagg caccaaggaa agcaaaatga actctctgac 420 acagcaggaa gggaaattaacctggttgat gctcacctta agagtgagca aacagaagca 480 agcaccaggc aaaattcccaagtactgcta tcagaaactg gaatttatga taatgaccct 540 gacctttgtt tcaggatgcaggaagggtct gaagtttatt ctaatccatg cctggaagaa 600 aacaaaccag gcattgtttatgcttccctg aaccattctg tcattggact gaactcaaga 660 ctggcaagaa atgtaaaagaagcaccaaca gaatatgcat ccatatgtgt gaggagt 717 41 777 DNA Homo sapiens 41aaagaatcat gtgatgtaca gctttatata aagagacaat ctgaacactc catcttagca 60ggagatccct ttgaactaga atgccctgtg aaatactgtg ctaacaggcc tcatgtgact 120tggtgcaagc tcaatggaac aacatgtgta aaacttgaag atagacaaac aagttggaag 180gaagagaaga acatttcatt tttcattcta cattttgaac cagtgcttcc taatgacaat 240gggtcatacc gctgttctgc aaattttcag tctaatctca ttgaaagcca ctcaacaact 300ctttatgtga cagatgtaaa aagtgcctca gaacgaccct ccaaggacga aatggcaagc 360agaccctggc tcctgtatag tttacttcct ttggggggat tgcctctact catcactacc 420tgtttctgcc tgttctgctg cctgagaagg caccaaggaa agcaaaatga actctctgac 480acagcaggaa gggaaattaa cctggttgat gctcacctta agagtgagca aacagaagca 540agcaccaggc aaaattccca agtactgcta tcagaaactg gaatttatga taatgaccct 600gacctttgtt tcaggatgca ggaagggtct gaagtttatt ctaatccatg cctggaagaa 660aacaaaccag gcattgttta tgcttccctg aaccattctg tcattggact gaactcaaga 720ctggcaagaa atgtaaaaga agcaccaaca gaatatgcat ccatatgtgt gaggagt 777 42870 DNA Homo sapiens 42 atgaagacat tgcctgccat gcttggaact gggaaattattttgggtctt cttcttaatc 60 ccatatctgg acatctggaa catccatggg aaagaatcatgtgatgtaca gctttatata 120 aagagacaat ctgaacactc catcttagca ggagatccctttgaactaga atgccctgtg 180 aaatactgtg ctaacaggcc tcatgtgact tggtgcaagctcaatggaac aacatgtgta 240 aaacttgaag atagacaaac aagttggaag gaagagaagaacatttcatt tttcattcta 300 cattttgaac cagtgcttcc taatgacaat gggtcataccgctgttctgc aaattttcag 360 tctaatctca ttgaaagcca ctcaacaact ctttatgtgacagatgtaaa aagtgcctca 420 gaacgaccct ccaaggacga aatggcaagc agaccctggctcctgtatag tttacttcct 480 ttggggggat tgcctctact catcactacc tgtttctgcctgttctgctg cctgagaagg 540 caccaaggaa agcaaaatga actctctgac acagcaggaagggaaattaa cctggttgat 600 gctcacctta agagtgagca aacagaagca agcaccaggcaaaattccca agtactgcta 660 tcagaaactg gaatttatga taatgaccct gacctttgtttcaggatgca ggaagggtct 720 gaagtttatt ctaatccatg cctggaagaa aacaaaccaggcattgttta tgcttccctg 780 aaccattctg tcattggact gaactcaaga ctggcaagaaatgtaaaaga agcaccaaca 840 gaatatgcat ccatatgtgt gaggagttaa 870 43 29 PRTMus musculus 43 Met Lys Thr Val Pro Ala Met Leu Gly Thr Pro Arg Leu PheArg Glu 1 5 10 15 Phe Phe Ile Leu His Leu Gly Leu Trp Ser Ile Leu Cys 2025 44 27 PRT Mus musculus 44 Glu Lys Ala Thr Lys Arg Asn Asp Glu Glu CysPro Val Gln Leu Thr 1 5 10 15 Ile Thr Arg Asn Ser Lys Gln Ser Ala ArgThr 20 25 45 68 PRT Mus musculus 45 Gly Glu Leu Phe Lys Ile Gln Cys ProVal Lys Tyr Cys Val His Arg 1 5 10 15 Pro Asn Val Thr Trp Cys Lys HisAsn Gly Thr Ile Cys Val Pro Leu 20 25 30 Glu Val Ser Pro Gln Leu Tyr ThrSer Trp Glu Glu Asn Gln Ser Val 35 40 45 Pro Val Phe Val Leu His Phe LysPro Ile His Leu Ser Asp Asn Gly 50 55 60 Ser Tyr Ser Cys 65 46 56 PRTMus musculus 46 Ser Thr Asn Phe Asn Ser Gln Val Ile Asn Ser His Ser ValThr Ile 1 5 10 15 His Val Thr Glu Arg Thr Gln Asn Ser Ser Glu His ProLeu Ile Thr 20 25 30 Val Ser Asp Ile Pro Asp Ala Thr Asn Ala Ser Gly ProSer Thr Met 35 40 45 Glu Glu Arg Pro Gly Arg Thr Trp 50 55 47 24 PRT Musmusculus 47 Leu Leu Tyr Thr Leu Leu Pro Leu Gly Ala Leu Leu Leu Leu LeuAla 1 5 10 15 Cys Val Cys Leu Leu Cys Phe Leu 20 48 37 PRT Mus musculus48 Lys Arg Ile Gln Gly Lys Glu Lys Lys Pro Ser Asp Leu Ala Gly Arg 1 510 15 Asp Thr Asn Leu Val Asp Ile Pro Ala Ser Ser Arg Thr Asn His Gln 2025 30 Ala Leu Pro Ser Gly 35 49 8 PRT Mus musculus 49 Thr Gly Ile TyrAsp Asn Asp Pro 1 5 50 21 PRT Mus musculus 50 Trp Ser Ser Met Gln AspGlu Ser Glu Leu Thr Ile Ser Leu Gln Ser 1 5 10 15 Glu Arg Asn Asn Gln 2051 9 PRT Mus musculus 51 Gly Ile Val Tyr Ala Ser Leu Asn His 1 5 52 14PRT Mus musculus 52 Cys Val Ile Gly Arg Asn Pro Arg Gln Glu Asn Asn MetGln 1 5 10 53 13 PRT Mus musculus 53 Glu Ala Pro Thr Glu Tyr Ala Ser IleCys Val Arg Ser 1 5 10 54 151 PRT Mus musculus 54 Glu Lys Ala Thr LysArg Asn Asp Glu Glu Cys Pro Val Gln Leu Thr 1 5 10 15 Ile Thr Arg AsnSer Lys Gln Ser Ala Arg Thr Gly Glu Leu Phe Lys 20 25 30 Ile Gln Cys ProVal Lys Tyr Cys Val His Arg Pro Asn Val Thr Trp 35 40 45 Cys Lys His AsnGly Thr Ile Cys Val Pro Leu Glu Val Ser Pro Gln 50 55 60 Leu Tyr Thr SerTrp Glu Glu Asn Gln Ser Val Pro Val Phe Val Leu 65 70 75 80 His Phe LysPro Ile His Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser 85 90 95 Thr Asn PheAsn Ser Gln Val Ile Asn Ser His Ser Val Thr Ile His 100 105 110 Val ThrGlu Arg Thr Gln Asn Ser Ser Glu His Pro Leu Ile Thr Val 115 120 125 SerAsp Ile Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met Glu 130 135 140Glu Arg Pro Gly Arg Thr Trp 145 150 55 180 PRT Mus musculus 55 Met LysThr Val Pro Ala Met Leu Gly Thr Pro Arg Leu Phe Arg Glu 1 5 10 15 PhePhe Ile Leu His Leu Gly Leu Trp Ser Ile Leu Cys Glu Lys Ala 20 25 30 ThrLys Arg Asn Asp Glu Glu Cys Pro Val Gln Leu Thr Ile Thr Arg 35 40 45 AsnSer Lys Gln Ser Ala Arg Thr Gly Glu Leu Phe Lys Ile Gln Cys 50 55 60 ProVal Lys Tyr Cys Val His Arg Pro Asn Val Thr Trp Cys Lys His 65 70 75 80Asn Gly Thr Ile Cys Val Pro Leu Glu Val Ser Pro Gln Leu Tyr Thr 85 90 95Ser Trp Glu Glu Asn Gln Ser Val Pro Val Phe Val Leu His Phe Lys 100 105110 Pro Ile His Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser Thr Asn Phe 115120 125 Asn Ser Gln Val Ile Asn Ser His Ser Val Thr Ile His Val Thr Glu130 135 140 Arg Thr Gln Asn Ser Ser Glu His Pro Leu Ile Thr Val Ser AspIle 145 150 155 160 Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met GluGlu Arg Pro 165 170 175 Gly Arg Thr Trp 180 56 175 PRT Mus musculus 56Glu Lys Ala Thr Lys Arg Asn Asp Glu Glu Cys Pro Val Gln Leu Thr 1 5 1015 Ile Thr Arg Asn Ser Lys Gln Ser Ala Arg Thr Gly Glu Leu Phe Lys 20 2530 Ile Gln Cys Pro Val Lys Tyr Cys Val His Arg Pro Asn Val Thr Trp 35 4045 Cys Lys His Asn Gly Thr Ile Cys Val Pro Leu Glu Val Ser Pro Gln 50 5560 Leu Tyr Thr Ser Trp Glu Glu Asn Gln Ser Val Pro Val Phe Val Leu 65 7075 80 His Phe Lys Pro Ile His Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser 8590 95 Thr Asn Phe Asn Ser Gln Val Ile Asn Ser His Ser Val Thr Ile His100 105 110 Val Thr Glu Arg Thr Gln Asn Ser Ser Glu His Pro Leu Ile ThrVal 115 120 125 Ser Asp Ile Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser ThrMet Glu 130 135 140 Glu Arg Pro Gly Arg Thr Trp Leu Leu Tyr Thr Leu LeuPro Leu Gly 145 150 155 160 Ala Leu Leu Leu Leu Leu Ala Cys Val Cys LeuLeu Cys Phe Leu 165 170 175 57 204 PRT Mus musculus 57 Met Lys Thr ValPro Ala Met Leu Gly Thr Pro Arg Leu Phe Arg Glu 1 5 10 15 Phe Phe IleLeu His Leu Gly Leu Trp Ser Ile Leu Cys Glu Lys Ala 20 25 30 Thr Lys ArgAsn Asp Glu Glu Cys Pro Val Gln Leu Thr Ile Thr Arg 35 40 45 Asn Ser LysGln Ser Ala Arg Thr Gly Glu Leu Phe Lys Ile Gln Cys 50 55 60 Pro Val LysTyr Cys Val His Arg Pro Asn Val Thr Trp Cys Lys His 65 70 75 80 Asn GlyThr Ile Cys Val Pro Leu Glu Val Ser Pro Gln Leu Tyr Thr 85 90 95 Ser TrpGlu Glu Asn Gln Ser Val Pro Val Phe Val Leu His Phe Lys 100 105 110 ProIle His Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser Thr Asn Phe 115 120 125Asn Ser Gln Val Ile Asn Ser His Ser Val Thr Ile His Val Thr Glu 130 135140 Arg Thr Gln Asn Ser Ser Glu His Pro Leu Ile Thr Val Ser Asp Ile 145150 155 160 Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met Glu Glu ArgPro 165 170 175 Gly Arg Thr Trp Leu Leu Tyr Thr Leu Leu Pro Leu Gly AlaLeu Leu 180 185 190 Leu Leu Leu Ala Cys Val Cys Leu Leu Cys Phe Leu 195200 58 65 PRT Mus musculus 58 Thr Gly Ile Tyr Asp Asn Asp Pro Trp SerSer Met Gln Asp Glu Ser 1 5 10 15 Glu Leu Thr Ile Ser Leu Gln Ser GluArg Asn Asn Gln Gly Ile Val 20 25 30 Tyr Ala Ser Leu Asn His Cys Val IleGly Arg Asn Pro Arg Gln Glu 35 40 45 Asn Asn Met Gln Glu Ala Pro Thr GluTyr Ala Ser Ile Cys Val Arg 50 55 60 Ser 65 59 102 PRT Mus musculus 59Lys Arg Ile Gln Gly Lys Glu Lys Lys Pro Ser Asp Leu Ala Gly Arg 1 5 1015 Asp Thr Asn Leu Val Asp Ile Pro Ala Ser Ser Arg Thr Asn His Gln 20 2530 Ala Leu Pro Ser Gly Thr Gly Ile Tyr Asp Asn Asp Pro Trp Ser Ser 35 4045 Met Gln Asp Glu Ser Glu Leu Thr Ile Ser Leu Gln Ser Glu Arg Asn 50 5560 Asn Gln Gly Ile Val Tyr Ala Ser Leu Asn His Cys Val Ile Gly Arg 65 7075 80 Asn Pro Arg Gln Glu Asn Asn Met Gln Glu Ala Pro Thr Glu Tyr Ala 8590 95 Ser Ile Cys Val Arg Ser 100 60 126 PRT Mus musculus 60 Leu Leu TyrThr Leu Leu Pro Leu Gly Ala Leu Leu Leu Leu Leu Ala 1 5 10 15 Cys ValCys Leu Leu Cys Phe Leu Lys Arg Ile Gln Gly Lys Glu Lys 20 25 30 Lys ProSer Asp Leu Ala Gly Arg Asp Thr Asn Leu Val Asp Ile Pro 35 40 45 Ala SerSer Arg Thr Asn His Gln Ala Leu Pro Ser Gly Thr Gly Ile 50 55 60 Tyr AspAsn Asp Pro Trp Ser Ser Met Gln Asp Glu Ser Glu Leu Thr 65 70 75 80 IleSer Leu Gln Ser Glu Arg Asn Asn Gln Gly Ile Val Tyr Ala Ser 85 90 95 LeuAsn His Cys Val Ile Gly Arg Asn Pro Arg Gln Glu Asn Asn Met 100 105 110Gln Glu Ala Pro Thr Glu Tyr Ala Ser Ile Cys Val Arg Ser 115 120 125 61250 PRT Mus musculus 61 Gly Glu Leu Phe Lys Ile Gln Cys Pro Val Lys TyrCys Val His Arg 1 5 10 15 Pro Asn Val Thr Trp Cys Lys His Asn Gly ThrIle Cys Val Pro Leu 20 25 30 Glu Val Ser Pro Gln Leu Tyr Thr Ser Trp GluGlu Asn Gln Ser Val 35 40 45 Pro Val Phe Val Leu His Phe Lys Pro Ile HisLeu Ser Asp Asn Gly 50 55 60 Ser Tyr Ser Cys Ser Thr Asn Phe Asn Ser GlnVal Ile Asn Ser His 65 70 75 80 Ser Val Thr Ile His Val Thr Glu Arg ThrGln Asn Ser Ser Glu His 85 90 95 Pro Leu Ile Thr Val Ser Asp Ile Pro AspAla Thr Asn Ala Ser Gly 100 105 110 Pro Ser Thr Met Glu Glu Arg Pro GlyArg Thr Trp Leu Leu Tyr Thr 115 120 125 Leu Leu Pro Leu Gly Ala Leu LeuLeu Leu Leu Ala Cys Val Cys Leu 130 135 140 Leu Cys Phe Leu Lys Arg IleGln Gly Lys Glu Lys Lys Pro Ser Asp 145 150 155 160 Leu Ala Gly Arg AspThr Asn Leu Val Asp Ile Pro Ala Ser Ser Arg 165 170 175 Thr Asn His GlnAla Leu Pro Ser Gly Thr Gly Ile Tyr Asp Asn Asp 180 185 190 Pro Trp SerSer Met Gln Asp Glu Ser Glu Leu Thr Ile Ser Leu Gln 195 200 205 Ser GluArg Asn Asn Gln Gly Ile Val Tyr Ala Ser Leu Asn His Cys 210 215 220 ValIle Gly Arg Asn Pro Arg Gln Glu Asn Asn Met Gln Glu Ala Pro 225 230 235240 Thr Glu Tyr Ala Ser Ile Cys Val Arg Ser 245 250 62 277 PRT Musmusculus 62 Glu Lys Ala Thr Lys Arg Asn Asp Glu Glu Cys Pro Val Gln LeuThr 1 5 10 15 Ile Thr Arg Asn Ser Lys Gln Ser Ala Arg Thr Gly Glu LeuPhe Lys 20 25 30 Ile Gln Cys Pro Val Lys Tyr Cys Val His Arg Pro Asn ValThr Trp 35 40 45 Cys Lys His Asn Gly Thr Ile Cys Val Pro Leu Glu Val SerPro Gln 50 55 60 Leu Tyr Thr Ser Trp Glu Glu Asn Gln Ser Val Pro Val PheVal Leu 65 70 75 80 His Phe Lys Pro Ile His Leu Ser Asp Asn Gly Ser TyrSer Cys Ser 85 90 95 Thr Asn Phe Asn Ser Gln Val Ile Asn Ser His Ser ValThr Ile His 100 105 110 Val Thr Glu Arg Thr Gln Asn Ser Ser Glu His ProLeu Ile Thr Val 115 120 125 Ser Asp Ile Pro Asp Ala Thr Asn Ala Ser GlyPro Ser Thr Met Glu 130 135 140 Glu Arg Pro Gly Arg Thr Trp Leu Leu TyrThr Leu Leu Pro Leu Gly 145 150 155 160 Ala Leu Leu Leu Leu Leu Ala CysVal Cys Leu Leu Cys Phe Leu Lys 165 170 175 Arg Ile Gln Gly Lys Glu LysLys Pro Ser Asp Leu Ala Gly Arg Asp 180 185 190 Thr Asn Leu Val Asp IlePro Ala Ser Ser Arg Thr Asn His Gln Ala 195 200 205 Leu Pro Ser Gly ThrGly Ile Tyr Asp Asn Asp Pro Trp Ser Ser Met 210 215 220 Gln Asp Glu SerGlu Leu Thr Ile Ser Leu Gln Ser Glu Arg Asn Asn 225 230 235 240 Gln GlyIle Val Tyr Ala Ser Leu Asn His Cys Val Ile Gly Arg Asn 245 250 255 ProArg Gln Glu Asn Asn Met Gln Glu Ala Pro Thr Glu Tyr Ala Ser 260 265 270Ile Cys Val Arg Ser 275 63 306 PRT Mus musculus 63 Met Lys Thr Val ProAla Met Leu Gly Thr Pro Arg Leu Phe Arg Glu 1 5 10 15 Phe Phe Ile LeuHis Leu Gly Leu Trp Ser Ile Leu Cys Glu Lys Ala 20 25 30 Thr Lys Arg AsnAsp Glu Glu Cys Pro Val Gln Leu Thr Ile Thr Arg 35 40 45 Asn Ser Lys GlnSer Ala Arg Thr Gly Glu Leu Phe Lys Ile Gln Cys 50 55 60 Pro Val Lys TyrCys Val His Arg Pro Asn Val Thr Trp Cys Lys His 65 70 75 80 Asn Gly ThrIle Cys Val Pro Leu Glu Val Ser Pro Gln Leu Tyr Thr 85 90 95 Ser Trp GluGlu Asn Gln Ser Val Pro Val Phe Val Leu His Phe Lys 100 105 110 Pro IleHis Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser Thr Asn Phe 115 120 125 AsnSer Gln Val Ile Asn Ser His Ser Val Thr Ile His Val Thr Glu 130 135 140Arg Thr Gln Asn Ser Ser Glu His Pro Leu Ile Thr Val Ser Asp Ile 145 150155 160 Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met Glu Glu Arg Pro165 170 175 Gly Arg Thr Trp Leu Leu Tyr Thr Leu Leu Pro Leu Gly Ala LeuLeu 180 185 190 Leu Leu Leu Ala Cys Val Cys Leu Leu Cys Phe Leu Lys ArgIle Gln 195 200 205 Gly Lys Glu Lys Lys Pro Ser Asp Leu Ala Gly Arg AspThr Asn Leu 210 215 220 Val Asp Ile Pro Ala Ser Ser Arg Thr Asn His GlnAla Leu Pro Ser 225 230 235 240 Gly Thr Gly Ile Tyr Asp Asn Asp Pro TrpSer Ser Met Gln Asp Glu 245 250 255 Ser Glu Leu Thr Ile Ser Leu Gln SerGlu Arg Asn Asn Gln Gly Ile 260 265 270 Val Tyr Ala Ser Leu Asn His CysVal Ile Gly Arg Asn Pro Arg Gln 275 280 285 Glu Asn Asn Met Gln Glu AlaPro Thr Glu Tyr Ala Ser Ile Cys Val 290 295 300 Arg Ser 305 64 87 DNAMus musculus 64 atgaagacag tgcctgccat gcttgggact cctcggttat ttagggaattcttcatcctc 60 catctgggcc tctggagcat cctttgt 87 65 81 DNA Mus musculus 65gagaaagcta ctaagaggaa tgatgaagag tgtccagtgc aacttactat tacgaggaat 60tccaaacagt ctgccaggac a 81 66 210 DNA Mus musculus 66 ggagagttatttaaaattca atgtcctgtg aaatactgtg ttcatagacc taatgtgact 60 tggtgtaagcacaatggaac aatctgtgta ccccttgagg ttagccctca gctatacact 120 agttgggaagaaaatcaatc agttccggtt tttgttctcc actttaaacc aatacatctc 180 agtgataatgggtcgtatag ctgttctaca 210 67 162 DNA Mus musculus 67 aacttcaattctcaagttat taatagccat tcagtaacca tccatgtgac agaaaggact 60 caaaactcttcagaacaccc actaataaca gtatctgaca tcccagatgc caccaatgcc 120 tcaggaccatccaccatgga agagaggcca ggcaggactt gg 162 68 72 DNA Mus musculus 68ctgctttaca ccttgcttcc tttgggggca ttgcttctgc tccttgcctg tgtctgcctg 60ctctgctttc tg 72 69 111 DNA Mus musculus 69 aaaaggatcc aagggaaagaaaagaagcct tctgacttgg caggaaggga cactaacctg 60 gttgatattc cagccagttccaggacaaat caccaagcac tgccatcagg a 111 70 24 DNA Mus musculus 70actggaattt atgataatga tccc 24 71 63 DNA Mus musculus 71 tggtctagcatgcaggatga atctgaattg acaattagct tgcaatcaga gagaaacaac 60 cag 63 72 27DNA Mus musculus 72 ggcattgttt atgcttcttt gaaccat 27 73 42 DNA Musmusculus 73 tgtgttattg gaaggaatcc aagacaggaa aacaacatgc ag 42 74 39 DNAMus musculus 74 gaggcaccca cagaatatgc atccatttgt gtgagaagt 39 75 453 DNAMus musculus 75 gagaaagcta ctaagaggaa tgatgaagag tgtccagtgc aacttactattacgaggaat 60 tccaaacagt ctgccaggac aggagagtta tttaaaattc aatgtcctgtgaaatactgt 120 gttcatagac ctaatgtgac ttggtgtaag cacaatggaa caatctgtgtaccccttgag 180 gttagccctc agctatacac tagttgggaa gaaaatcaat cagttccggtttttgttctc 240 cactttaaac caatacatct cagtgataat gggtcgtata gctgttctacaaacttcaat 300 tctcaagtta ttaatagcca ttcagtaacc atccatgtga cagaaaggactcaaaactct 360 tcagaacacc cactaataac agtatctgac atcccagatg ccaccaatgcctcaggacca 420 tccaccatgg aagagaggcc aggcaggact tgg 453 76 540 DNA Musmusculus 76 atgaagacag tgcctgccat gcttgggact cctcggttat ttagggaattcttcatcctc 60 catctgggcc tctggagcat cctttgtgag aaagctacta agaggaatgatgaagagtgt 120 ccagtgcaac ttactattac gaggaattcc aaacagtctg ccaggacaggagagttattt 180 aaaattcaat gtcctgtgaa atactgtgtt catagaccta atgtgacttggtgtaagcac 240 aatggaacaa tctgtgtacc ccttgaggtt agccctcagc tatacactagttgggaagaa 300 aatcaatcag ttccggtttt tgttctccac tttaaaccaa tacatctcagtgataatggg 360 tcgtatagct gttctacaaa cttcaattct caagttatta atagccattcagtaaccatc 420 catgtgacag aaaggactca aaactcttca gaacacccac taataacagtatctgacatc 480 ccagatgcca ccaatgcctc aggaccatcc accatggaag agaggccaggcaggacttgg 540 77 525 DNA Mus musculus 77 gagaaagcta ctaagaggaatgatgaagag tgtccagtgc aacttactat tacgaggaat 60 tccaaacagt ctgccaggacaggagagtta tttaaaattc aatgtcctgt gaaatactgt 120 gttcatagac ctaatgtgacttggtgtaag cacaatggaa caatctgtgt accccttgag 180 gttagccctc agctatacactagttgggaa gaaaatcaat cagttccggt ttttgttctc 240 cactttaaac caatacatctcagtgataat gggtcgtata gctgttctac aaacttcaat 300 tctcaagtta ttaatagccattcagtaacc atccatgtga cagaaaggac tcaaaactct 360 tcagaacacc cactaataacagtatctgac atcccagatg ccaccaatgc ctcaggacca 420 tccaccatgg aagagaggccaggcaggact tggctgcttt acaccttgct tcctttgggg 480 gcattgcttc tgctccttgcctgtgtctgc ctgctctgct ttctg 525 78 612 DNA Mus musculus 78 atgaagacagtgcctgccat gcttgggact cctcggttat ttagggaatt cttcatcctc 60 catctgggcctctggagcat cctttgtgag aaagctacta agaggaatga tgaagagtgt 120 ccagtgcaacttactattac gaggaattcc aaacagtctg ccaggacagg agagttattt 180 aaaattcaatgtcctgtgaa atactgtgtt catagaccta atgtgacttg gtgtaagcac 240 aatggaacaatctgtgtacc ccttgaggtt agccctcagc tatacactag ttgggaagaa 300 aatcaatcagttccggtttt tgttctccac tttaaaccaa tacatctcag tgataatggg 360 tcgtatagctgttctacaaa cttcaattct caagttatta atagccattc agtaaccatc 420 catgtgacagaaaggactca aaactcttca gaacacccac taataacagt atctgacatc 480 ccagatgccaccaatgcctc aggaccatcc accatggaag agaggccagg caggacttgg 540 ctgctttacaccttgcttcc tttgggggca ttgcttctgc tccttgcctg tgtctgcctg 600 ctctgctttc tg612 79 195 DNA Mus musculus 79 actggaattt atgataatga tccctggtctagcatgcagg atgaatctga attgacaatt 60 agcttgcaat cagagagaaa caaccagggcattgtttatg cttctttgaa ccattgtgtt 120 attggaagga atccaagaca ggaaaacaacatgcaggagg cacccacaga atatgcatcc 180 atttgtgtga gaagt 195 80 306 DNA Musmusculus 80 aaaaggatcc aagggaaaga aaagaagcct tctgacttgg caggaagggacactaacctg 60 gttgatattc cagccagttc caggacaaat caccaagcac tgccatcaggaactggaatt 120 tatgataatg atccctggtc tagcatgcag gatgaatctg aattgacaattagcttgcaa 180 tcagagagaa acaaccaggg cattgtttat gcttctttga accattgtgttattggaagg 240 aatccaagac aggaaaacaa catgcaggag gcacccacag aatatgcatccatttgtgtg 300 agaagt 306 81 378 DNA Mus musculus 81 ctgctttacaccttgcttcc tttgggggca ttgcttctgc tccttgcctg tgtctgcctg 60 ctctgctttctgaaaaggat ccaagggaaa gaaaagaagc cttctgactt ggcaggaagg 120 gacactaacctggttgatat tccagccagt tccaggacaa atcaccaagc actgccatca 180 ggaactggaatttatgataa tgatccctgg tctagcatgc aggatgaatc tgaattgaca 240 attagcttgcaatcagagag aaacaaccag ggcattgttt atgcttcttt gaaccattgt 300 gttattggaaggaatccaag acaggaaaac aacatgcagg aggcacccac agaatatgca 360 tccatttgtgtgagaagt 378 82 750 DNA Mus musculus 82 ggagagttat ttaaaattca atgtcctgtgaaatactgtg ttcatagacc taatgtgact 60 tggtgtaagc acaatggaac aatctgtgtaccccttgagg ttagccctca gctatacact 120 agttgggaag aaaatcaatc agttccggtttttgttctcc actttaaacc aatacatctc 180 agtgataatg ggtcgtatag ctgttctacaaacttcaatt ctcaagttat taatagccat 240 tcagtaacca tccatgtgac agaaaggactcaaaactctt cagaacaccc actaataaca 300 gtatctgaca tcccagatgc caccaatgcctcaggaccat ccaccatgga agagaggcca 360 ggcaggactt ggctgcttta caccttgcttcctttggggg cattgcttct gctccttgcc 420 tgtgtctgcc tgctctgctt tctgaaaaggatccaaggga aagaaaagaa gccttctgac 480 ttggcaggaa gggacactaa cctggttgatattccagcca gttccaggac aaatcaccaa 540 gcactgccat caggaactgg aatttatgataatgatccct ggtctagcat gcaggatgaa 600 tctgaattga caattagctt gcaatcagagagaaacaacc agggcattgt ttatgcttct 660 ttgaaccatt gtgttattgg aaggaatccaagacaggaaa acaacatgca ggaggcaccc 720 acagaatatg catccatttg tgtgagaagt750 83 831 DNA Mus musculus 83 gagaaagcta ctaagaggaa tgatgaagagtgtccagtgc aacttactat tacgaggaat 60 tccaaacagt ctgccaggac aggagagttatttaaaattc aatgtcctgt gaaatactgt 120 gttcatagac ctaatgtgac ttggtgtaagcacaatggaa caatctgtgt accccttgag 180 gttagccctc agctatacac tagttgggaagaaaatcaat cagttccggt ttttgttctc 240 cactttaaac caatacatct cagtgataatgggtcgtata gctgttctac aaacttcaat 300 tctcaagtta ttaatagcca ttcagtaaccatccatgtga cagaaaggac tcaaaactct 360 tcagaacacc cactaataac agtatctgacatcccagatg ccaccaatgc ctcaggacca 420 tccaccatgg aagagaggcc aggcaggacttggctgcttt acaccttgct tcctttgggg 480 gcattgcttc tgctccttgc ctgtgtctgcctgctctgct ttctgaaaag gatccaaggg 540 aaagaaaaga agccttctga cttggcaggaagggacacta acctggttga tattccagcc 600 agttccagga caaatcacca agcactgccatcaggaactg gaatttatga taatgatccc 660 tggtctagca tgcaggatga atctgaattgacaattagct tgcaatcaga gagaaacaac 720 cagggcattg tttatgcttc tttgaaccattgtgttattg gaaggaatcc aagacaggaa 780 aacaacatgc aggaggcacc cacagaatatgcatccattt gtgtgagaag t 831 84 921 DNA Mus musculus 84 atgaagacagtgcctgccat gcttgggact cctcggttat ttagggaatt cttcatcctc 60 catctgggcctctggagcat cctttgtgag aaagctacta agaggaatga tgaagagtgt 120 ccagtgcaacttactattac gaggaattcc aaacagtctg ccaggacagg agagttattt 180 aaaattcaatgtcctgtgaa atactgtgtt catagaccta atgtgacttg gtgtaagcac 240 aatggaacaatctgtgtacc ccttgaggtt agccctcagc tatacactag ttgggaagaa 300 aatcaatcagttccggtttt tgttctccac tttaaaccaa tacatctcag tgataatggg 360 tcgtatagctgttctacaaa cttcaattct caagttatta atagccattc agtaaccatc 420 catgtgacagaaaggactca aaactcttca gaacacccac taataacagt atctgacatc 480 ccagatgccaccaatgcctc aggaccatcc accatggaag agaggccagg caggacttgg 540 ctgctttacaccttgcttcc tttgggggca ttgcttctgc tccttgcctg tgtctgcctg 600 ctctgctttctgaaaaggat ccaagggaaa gaaaagaagc cttctgactt ggcaggaagg 660 gacactaacctggttgatat tccagccagt tccaggacaa atcaccaagc actgccatca 720 ggaactggaatttatgataa tgatccctgg tctagcatgc aggatgaatc tgaattgaca 780 attagcttgcaatcagagag aaacaaccag ggcattgttt atgcttcttt gaaccattgt 840 gttattggaaggaatccaag acaggaaaac aacatgcagg aggcacccac agaatatgca 900 tccatttgtgtgagaagtta a 921 85 179 PRT Mus musculus 85 Met Lys Thr Val Pro Ala MetLeu Gly Thr Pro Arg Leu Phe Arg Glu 1 5 10 15 Phe Phe Ile Leu His LeuGly Leu Trp Ser Ile Leu Cys Glu Lys Ala 20 25 30 Thr Lys Arg Asn Asp GluGlu Cys Glu Val Gln Leu Asn Ile Lys Arg 35 40 45 Asn Ser Lys His Ser AlaTrp Thr Gly Glu Leu Phe Lys Ile Glu Cys 50 55 60 Pro Val Lys Tyr Cys ValHis Arg Pro His Val Thr Trp Cys Lys His 65 70 75 80 Asn Gly Thr Ile TrpVal Pro Leu Glu Val Gly Pro Gln Leu Tyr Thr 85 90 95 Ser Trp Glu Glu AsnArg Ser Val Pro Val Phe Val Leu His Phe Lys 100 105 110 Pro Ile His LeuSer Asp Asn Gly Ser Tyr Ser Cys Ser Thr Asn Phe 115 120 125 Asn Ser GlnVal Ile Asn Ser His Ser Val Thr Ile His Val Arg Glu 130 135 140 Arg ThrGln Asn Ser Ser Glu His Pro Leu Ile Thr Val Ser Asp Ile 145 150 155 160Pro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met Glu Glu Arg Pro 165 170175 Gly Arg Thr 86 150 PRT Mus musculus 86 Glu Lys Ala Thr Lys Arg AsnAsp Glu Glu Cys Glu Val Gln Leu Asn 1 5 10 15 Ile Lys Arg Asn Ser LysHis Ser Ala Trp Thr Gly Glu Leu Phe Lys 20 25 30 Ile Glu Cys Pro Val LysTyr Cys Val His Arg Pro His Val Thr Trp 35 40 45 Cys Lys His Asn Gly ThrIle Trp Val Pro Leu Glu Val Gly Pro Gln 50 55 60 Leu Tyr Thr Ser Trp GluGlu Asn Arg Ser Val Pro Val Phe Val Leu 65 70 75 80 His Phe Lys Pro IleHis Leu Ser Asp Asn Gly Ser Tyr Ser Cys Ser 85 90 95 Thr Asn Phe Asn SerGln Val Ile Asn Ser His Ser Val Thr Ile His 100 105 110 Val Arg Glu ArgThr Gln Asn Ser Ser Glu His Pro Leu Ile Thr Val 115 120 125 Ser Asp IlePro Asp Ala Thr Asn Ala Ser Gly Pro Ser Thr Met Glu 130 135 140 Glu ArgPro Gly Arg Thr 145 150 87 29 PRT Mus musculus 87 Met Lys Thr Val ProAla Met Leu Gly Thr Pro Arg Leu Phe Arg Glu 1 5 10 15 Phe Phe Ile LeuHis Leu Gly Leu Trp Ser Ile Leu Cys 20 25 88 68 PRT Mus musculus 88 GlyGlu Leu Phe Lys Ile Glu Cys Pro Val Lys Tyr Cys Val His Arg 1 5 10 15Pro His Val Thr Trp Cys Lys His Asn Gly Thr Ile Trp Val Pro Leu 20 25 30Glu Val Gly Pro Gln Leu Tyr Thr Ser Trp Glu Glu Asn Arg Ser Val 35 40 45Pro Val Phe Val Leu His Phe Lys Pro Ile His Leu Ser Asp Asn Gly 50 55 60Ser Tyr Ser Cys 65 89 537 DNA Mus musculus 89 atgaagacag tgcctgccatgcttgggact cctcggttat ttagggaatt cttcatcctc 60 catctgggcc tctggagcatcctttgtgag aaagctacta agaggaatga tgaagagtgt 120 gaagtgcaac ttaatattaagaggaattcc aaacactctg cctggacagg agagttattt 180 aaaattgaat gtcctgtgaaatactgtgtt catagacctc atgtgacttg gtgtaagcac 240 aatggaacaa tctgggtaccccttgaagtt ggtcctcagc tatacactag ttgggaagaa 300 aatcgatcag ttccggtttttgttctccat tttaaaccaa tacatctcag tgataacggg 360 tcgtatagct gttctacaaacttcaattct caagttatta atagccattc agtaaccatc 420 catgtgagag aaaggactcaaaactcttca gaacacccac taataacagt atctgacatc 480 ccagatgcca ccaatgcctcaggaccatcc accatggaag agaggccagg caggact 537 90 450 DNA Mus musculus 90gagaaagcta ctaagaggaa tgatgaagag tgtgaagtgc aacttaatat taagaggaat 60tccaaacact ctgcctggac aggagagtta tttaaaattg aatgtcctgt gaaatactgt 120gttcatagac ctcatgtgac ttggtgtaag cacaatggaa caatctgggt accccttgaa 180gttggtcctc agctatacac tagttgggaa gaaaatcgat cagttccggt ttttgttctc 240cattttaaac caatacatct cagtgataac gggtcgtata gctgttctac aaacttcaat 300tctcaagtta ttaatagcca ttcagtaacc atccatgtga gagaaaggac tcaaaactct 360tcagaacacc cactaataac agtatctgac atcccagatg ccaccaatgc ctcaggacca 420tccaccatgg aagagaggcc aggcaggact 450 91 87 DNA Mus musculus 91atgaagacag tgcctgccat gcttgggact cctcggttat ttagggaatt cttcatcctc 60catctgggcc tctggagcat cctttgt 87 92 204 DNA Mus musculus 92 ggagagttatttaaaattga atgtcctgtg aaatactgtg ttcatagacc tcatgtgact 60 tggtgtaagcacaatggaac aatctgggta ccccttgaag ttggtcctca gctatacact 120 agttgggaagaaaatcgatc agttccggtt tttgttctcc attttaaacc aatacatctc 180 agtgataacgggtcgtatag ctgt 204 93 6 PRT Artificial Sequence Xaa = any amino acid 93Ile Xaa Tyr Xaa Xaa Leu 1 5 94 6 PRT Artificial Sequence Synthesized 94Ile Val Tyr Ala Ser Leu 1 5 95 6 PRT Artificial Sequence Xaa = any aminoacid 95 Thr Xaa Tyr Xaa Xaa Ile 1 5 96 6 PRT Artificial SequenceSynthesized 96 Thr Glu Tyr Ala Ser Ile 1 5 97 232 PRT Homo sapiens 97Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 1015 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 20 2530 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 4045 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 5560 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 7075 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 8590 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly GlnPro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp GluLeu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly PheTyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly GlnPro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser AspGly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser ArgTrp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu AlaLeu His Asn His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro GlyLys 225 230 98 521 PRT Artificial Sequence Fusion polypeptide 98 Glu ProLys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 ProGlu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 20 25 30 LysAsp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 ValAsp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 AspGly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr130 135 140 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr ProSer 145 150 155 160 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro GluAsn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly SerPhe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp GlnGln Gly Asn Val Phe 195 200 205 Ser Cys Ser Val Met His Glu Ala Leu HisAsn His Tyr Thr Gln Lys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys MetLys Thr Leu Pro Ala Met Leu 225 230 235 240 Gly Thr Gly Lys Leu Phe TrpVal Phe Phe Leu Ile Pro Tyr Leu Asp 245 250 255 Ile Trp Asn Ile His GlyLys Glu Ser Cys Asp Val Gln Leu Tyr Ile 260 265 270 Lys Arg Gln Ser GluHis Ser Ile Leu Ala Gly Asp Pro Phe Glu Leu 275 280 285 Glu Cys Pro ValLys Tyr Cys Ala Asn Arg Pro His Val Thr Trp Cys 290 295 300 Lys Leu AsnGly Thr Thr Cys Val Lys Leu Glu Asp Arg Gln Thr Ser 305 310 315 320 TrpLys Glu Glu Lys Asn Ile Ser Phe Phe Ile Leu His Phe Glu Pro 325 330 335Val Leu Pro Asn Asp Asn Gly Ser Tyr Arg Cys Ser Ala Asn Phe Gln 340 345350 Ser Asn Leu Ile Glu Ser His Ser Thr Thr Leu Tyr Val Thr Asp Val 355360 365 Lys Ser Ala Ser Glu Arg Pro Ser Lys Asp Glu Met Ala Ser Arg Pro370 375 380 Trp Leu Leu Tyr Ser Leu Leu Pro Leu Gly Gly Leu Pro Leu LeuIle 385 390 395 400 Thr Thr Cys Phe Cys Leu Phe Cys Cys Leu Arg Arg HisGln Gly Lys 405 410 415 Gln Asn Glu Leu Ser Asp Thr Ala Gly Arg Glu IleAsn Leu Val Asp 420 425 430 Ala His Leu Lys Ser Glu Gln Thr Glu Ala SerThr Arg Gln Asn Ser 435 440 445 Gln Val Leu Leu Ser Glu Thr Gly Ile TyrAsp Asn Asp Pro Asp Leu 450 455 460 Cys Phe Arg Met Gln Glu Gly Ser GluVal Tyr Ser Asn Pro Cys Leu 465 470 475 480 Glu Glu Asn Lys Pro Gly IleVal Tyr Ala Ser Leu Asn His Ser Val 485 490 495 Ile Gly Leu Asn Ser ArgLeu Ala Arg Asn Val Lys Glu Ala Pro Thr 500 505 510 Glu Tyr Ala Ser IleCys Val Arg Ser 515 520 99 538 PRT Artificial Sequence Fusionpolypeptide 99 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro CysPro Ala 1 5 10 15 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe ProPro Lys Pro 20 25 30 Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val ThrCys Val Val 35 40 45 Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe AsnTrp Tyr Val 50 55 60 Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro ArgGlu Glu Gln 65 70 75 80 Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu ThrVal Leu His Gln 85 90 95 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys ValSer Asn Lys Ala 100 105 110 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser LysAla Lys Gly Gln Pro 115 120 125 Arg Glu Pro Gln Val Tyr Thr Leu Pro ProSer Arg Asp Glu Leu Thr 130 135 140 Lys Asn Gln Val Ser Leu Thr Cys LeuVal Lys Gly Phe Tyr Pro Ser 145 150 155 160 Asp Ile Ala Val Glu Trp GluSer Asn Gly Gln Pro Glu Asn Asn Tyr 165 170 175 Lys Thr Thr Pro Pro ValLeu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr ValAsp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 195 200 205 Ser Cys Ser ValMet His Glu Ala Leu His Asn His Tyr Thr Gln Lys 210 215 220 Ser Leu SerLeu Ser Pro Gly Lys Met Lys Thr Val Pro Ala Met Leu 225 230 235 240 GlyThr Pro Arg Leu Phe Arg Glu Phe Phe Ile Leu His Leu Gly Leu 245 250 255Trp Ser Ile Leu Cys Glu Lys Ala Thr Lys Arg Asn Asp Glu Glu Cys 260 265270 Pro Val Gln Leu Thr Ile Thr Arg Asn Ser Lys Gln Ser Ala Arg Thr 275280 285 Gly Glu Leu Phe Lys Ile Gln Cys Pro Val Lys Tyr Cys Val His Arg290 295 300 Pro Asn Val Thr Trp Cys Lys His Asn Gly Thr Ile Cys Val ProLeu 305 310 315 320 Glu Val Ser Pro Gln Leu Tyr Thr Ser Trp Glu Glu AsnGln Ser Val 325 330 335 Pro Val Phe Val Leu His Phe Lys Pro Ile His LeuSer Asp Asn Gly 340 345 350 Ser Tyr Ser Cys Ser Thr Asn Phe Asn Ser GlnVal Ile Asn Ser His 355 360 365 Ser Val Thr Ile His Val Thr Glu Arg ThrGln Asn Ser Ser Glu His 370 375 380 Pro Leu Ile Thr Val Ser Asp Ile ProAsp Ala Thr Asn Ala Ser Gly 385 390 395 400 Pro Ser Thr Met Glu Glu ArgPro Gly Arg Thr Trp Leu Leu Tyr Thr 405 410 415 Leu Leu Pro Leu Gly AlaLeu Leu Leu Leu Leu Ala Cys Val Cys Leu 420 425 430 Leu Cys Phe Leu LysArg Ile Gln Gly Lys Glu Lys Lys Pro Ser Asp 435 440 445 Leu Ala Gly ArgAsp Thr Asn Leu Val Asp Ile Pro Ala Ser Ser Arg 450 455 460 Thr Asn HisGln Ala Leu Pro Ser Gly Thr Gly Ile Tyr Asp Asn Asp 465 470 475 480 ProTrp Ser Ser Met Gln Asp Glu Ser Glu Leu Thr Ile Ser Leu Gln 485 490 495Ser Glu Arg Asn Asn Gln Gly Ile Val Tyr Ala Ser Leu Asn His Cys 500 505510 Val Ile Gly Arg Asn Pro Arg Gln Glu Asn Asn Met Gln Glu Ala Pro 515520 525 Thr Glu Tyr Ala Ser Ile Cys Val Arg Ser 530 535 100 411 PRTArtificial Sequence Fusion polypeptide 100 Glu Pro Lys Ser Cys Asp LysThr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Glu Leu Leu Gly GlyPro Ser Val Phe Leu Phe Pro Pro Lys Pro 20 25 30 Lys Asp Thr Leu Met IleSer Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45 Val Asp Val Ser His GluAsp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60 Asp Gly Val Glu Val HisAsn Ala Lys Thr Lys Pro Arg Glu Glu Gln 65 70 75 80 Tyr Asn Ser Thr TyrArg Val Val Ser Val Leu Thr Val Leu His Gln 85 90 95 Asp Trp Leu Asn GlyLys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110 Leu Pro Ala ProIle Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 115 120 125 Arg Glu ProGln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 130 135 140 Lys AsnGln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 165 170175 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe195 200 205 Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr GlnLys 210 215 220 Ser Leu Ser Leu Ser Pro Gly Lys Met Lys Thr Val Pro AlaMet Leu 225 230 235 240 Gly Thr Pro Arg Leu Phe Arg Glu Phe Phe Ile LeuHis Leu Gly Leu 245 250 255 Trp Ser Ile Leu Cys Glu Lys Ala Thr Lys ArgAsn Asp Glu Glu Cys 260 265 270 Glu Val Gln Leu Asn Ile Lys Arg Asn SerLys His Ser Ala Trp Thr 275 280 285 Gly Glu Leu Phe Lys Ile Glu Cys ProVal Lys Tyr Cys Val His Arg 290 295 300 Pro His Val Thr Trp Cys Lys HisAsn Gly Thr Ile Trp Val Pro Leu 305 310 315 320 Glu Val Gly Pro Gln LeuTyr Thr Ser Trp Glu Glu Asn Arg Ser Val 325 330 335 Pro Val Phe Val LeuHis Phe Lys Pro Ile His Leu Ser Asp Asn Gly 340 345 350 Ser Tyr Ser CysSer Thr Asn Phe Asn Ser Gln Val Ile Asn Ser His 355 360 365 Ser Val ThrIle His Val Arg Glu Arg Thr Gln Asn Ser Ser Glu His 370 375 380 Pro LeuIle Thr Val Ser Asp Ile Pro Asp Ala Thr Asn Ala Ser Gly 385 390 395 400Pro Ser Thr Met Glu Glu Arg Pro Gly Arg Thr 405 410 101 8 PRT ArtificialSequence Synthesized 101 Asp Tyr Lys Asp Asp Asp Asp Lys 1 5 102 10 PRTArtificial Sequence Synthesized 102 Glu Gln Lys Leu Ile Ser Glu Glu AspLeu 1 5 10 103 7 PRT Artificial Sequence Synthesized 103 Thr Gly Ile TyrAsp Asn Asp 1 5 104 990 DNA Homo sapiens 104 actggggtag gtaaactgacccaactctgc agcactcaga agacgaagca aagccttcta 60 cttgagcagt ttttccatcactgatatgtg caggaaatga agacattgcc tgccatgctt 120 ggaactggga aattattttgggtcttcttc ttaatcccat atctggacat ctggaacatc 180 catgggaaag aatcatgtgatgtacagctt tatataaaga gacaatctga acactccatc 240 ttagcaggag atccctttgaactagaatgc cctgtgaaat actgtgctaa caggcctcat 300 gtgacttggt gcaagctcaatggaacaaca tgtgtaaaac ttgaagatag acaaacaagt 360 tggaaggaag agaagaacatttcatttttc attctacatt ttgaaccagt gcttcctaat 420 gacaatgggt cataccgctgttctgcaaat tttcagtcta atctcattga aagccactca 480 acaactcttt atgtgacagatgtaaaaagt gcctcagaac gaccctccaa ggacgaaatg 540 gcaagcagac cctggctcctgtatagttta cttcctttgg ggggattgcc tctactcatc 600 actacctgtt tctgcctgttctgctgcctg agaaggcacc aaggaaagca aaatgaactc 660 tctgacacag caggaagggaaattaacctg gttgatgctc accttaagag tgagcaaaca 720 gaagcaagca ccaggcaaaattcccaagta ctgctatcag aaactggaat ttatgataat 780 gaccctgacc tttgtttcaggatgcaggaa gggtctgaag tttattctaa tccatgcctg 840 gaagaaaaca aaccaggcattgtttatgct tccctgaacc attctgtcat tggactgaac 900 tcaagactgg caagaaatgtaaaagaagca ccaacagaat atgcatccat atgtgtgagg 960 agttaagtct gttttctgacctcccaacag 990 105 1250 DNA Mus musculus 105 agatctctag ggaggaagaggaagtttgcc cttacctgac acgtgctggg aatgaagaca 60 gtgcctgcca tgcttgggactcctcggtta tttagggaat tcttcatcct ccatctgggc 120 ctctggagca tcctttgtgagaaagctact aagaggaatg atgaagagtg tccagtgcaa 180 cttactatta cgaggaattccaaacagtct gccaggacag gagagttatt taaaattcaa 240 tgtcctgtga aatactgtgttcatagacct aatgtgactt ggtgtaagca caatggaaca 300 atctgtgtac cccttgaggttagccctcag ctatacacta gttgggaaga aaatcaatca 360 gttccggttt ttgttctccactttaaacca atacatctca gtgataatgg gtcgtatagc 420 tgttctacaa acttcaattctcaagttatt aatagccatt cagtaaccat ccatgtgaca 480 gaaaggactc aaaactcttcagaacaccca ctaataacag tatctgacat cccagatgcc 540 accaatgcct caggaccatccaccatggaa gagaggccag gcaggacttg gctgctttac 600 accttgcttc ctttgggggcattgcttctg ctccttgcct gtgtctgcct gctctgcttt 660 ctgaaaagga tccaagggaaagaaaagaag ccttctgact tggcaggaag ggacactaac 720 ctggttgata ttccagccagttccaggaca aatcaccaag cactgccatc aggaactgga 780 atttatgata atgatccctggtctagcatg caggatgaat ctgaattgac aattagcttg 840 caatcagaga gaaacaaccagggcattgtt tatgcttctt tgaaccattg tgttattgga 900 aggaatccaa gacaggaaaacaacatgcag gaggcaccca cagaatatgc atccatttgt 960 gtgagaagtt aaacctgccactgagccagg cagcctacac tgcatgagtg cctgtcaata 1020 cctctgtctg gaccttcagtttcaaataac cttcaacctg gaaagtttca attaagatgc 1080 tctgtgctgg tgctgcgtcttaaaggtcca tgaagtattt agttaaaatt tctcctgaaa 1140 actttgggag agttttgtacaagacagact cttccctggg aaaactcata ttacgaatag 1200 cagaataata gggcttttaaaattgaccat gtcgcaccat gtcgtaccca 1250 106 957 DNA Mus musculus 106ggcccgggat ctatgaagac agtgcctgcc atgcttggga ctcctcggtt atttagggaa 60ttcttcatcc tccatctggg cctctggagc atcctttgtg agaaagctac taagaggaat 120gatgaagagt gtccagtgca acttactatt acgaggaatt ccaaacagtc tgccaggaca 180ggagagttat ttaaaattca atgtcctgtg aaatactgtg ttcatagacc taatgtgact 240tggtgtaagc acaatggaac aatctgtgta ccccttgagg ttagccctca gctatacact 300agttgggaag aaaatcaatc agttccggtt tttgttctcc actttaaacc aatacatctc 360agtgataatg ggtcgtatag ctgttctaca aacttcaatt ctcaagttat taatagccat 420tcagtaacca tccatgtgac agaaaggact caaaactctt cagaacaccc actaataaca 480gtatctgaca tcccagatgc caccaatgcc tcaggaccat ccaccatgga agagaggcca 540ggcaggactt ggctgcttta caccttgctt cctttggggg cattgcttct gctccttgcc 600tgtgtctgcc tgctctgctt tctgaaaagg atccaaggga aagaaaagaa gccttctgac 660ttggcaggaa gggacactaa cctggttgat attccagcca gttccaggac aaatcaccaa 720gcactgccat caggaactgg aatttatgat aatgatccct ggtctagcat gcaggatgaa 780tctgaattga caattagctt gcaatcagag agaaacaacc agggcattgt ttatgcttct 840ttgaaccatt gtgttattgg aaggaatcca agacaggaaa acaacatgca ggaggcaccc 900acagaatatg catccatttg tgtgagaagt gcagatccac cggtcgccac catggtg 957 1071722 DNA Mus musculus 107 ggatctcgag ctcaagcttc gaattctgca gtcgacggtaccgcgggccc gggatctatg 60 aagacagtgc ctgccatgct tgggactcct cggttatttagggaattctt catcctccat 120 ctgggcctct ggagcatcct ttgtgagaaa gctactaagaggaatgatga agagtgtcca 180 gtgcaactta ctattacgag gaattccaaa cagtctgccaggacaggaga gttatttaaa 240 attcaatgtc ctgtgaaata ctgtgttcat agacctaatgtgacttggtg taagcacaat 300 ggaacaatct gtgtacccct tgaggttagc cctcagctatacactagttg ggaagaaaat 360 caatcagttc cggtttttgt tctccacttt aaaccaatacatctcagtga taatgggtcg 420 tatagctgtt ctacaaactt caattctcaa gttattaatagccattcagt aaccatccat 480 gtgacagaaa ggactcaaaa ctcttcagaa cacccactaataacagtatc tgacatccca 540 gatgccacca atgcctcagg accatccacc atggaagagaggccaggcag gacttggctg 600 ctttacacct tgcttccttt gggggcattg cttctgctccttgcctgtgt ctgcctgctc 660 tgctttctga aaaggatcca agggaaagaa aagaagccttctgacttggc aggaagggac 720 actaacctgg ttgatattcc agccagttcc aggacaaatcaccaagcact gccatcagga 780 actggaattt atgataatga tccctggtct agcatgcaggatgaatctga attgacaatt 840 agcttgcaat cagagagaaa caaccagggc attgtttatgcttctttgaa ccattgtgtt 900 attggaagga atccaagaca ggaaaacaac atgcaggaggcacccacaga atatgcatcc 960 atttgtgtga gaagtgcaga tccaccggtc gccaccatggtgagcaaggg cgaggagctg 1020 ttcaccgggg tggtgcccat cctggtcgag ctggacggcgacgtaaacgg ccacaagttc 1080 agcgtgtccg gcgagggcga gggcgatgcc acctacggcaagctgaccct gaagttcatc 1140 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcgtgaccacctt cggctacggc 1200 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagcacgacttctt caagtccgcc 1260 atgcccgaag gctacgtcca ggagcgcacc atcttcttcaaggacgacgg caactacaag 1320 acccgcgccg aggtgaagtt cgagggcgac accctggtgaaccgcatcga gctgaagggc 1380 atcgacttca aggaggacgg caacatcctg gggcacaagctggagtacaa ctacaacagc 1440 cacaacgtct atatcatggc cgacaagcag aagaacggcatcaaggtgaa cttcaagatc 1500 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgaccactaccagca gaacaccccc 1560 atcggcgacg gccccgtgct gctgcccgac aaccactacctgagctacca gtccgccctg 1620 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgctggagttcgt gaccgccgcc 1680 gggatcactc tcggcatgga cgagctgtac aagtaaagcggc 1722 108 609 DNA Mus musculus 108 ggatccaagg gaaagaaaag aagccttctgacttggcagg aagggacact aacctggttg 60 atattccagc cagttccagg acaaatcaccaagcactgcc atcaggaact ggaatttatg 120 ataatgatcc ctggtctagc atgcaggatgaatctgaatt gacaattagc ttgcaatcag 180 agagaaacaa ccagggcatt gtttatgcttctttgaacca ttgtgttatt ggaaggaatc 240 caagacagga aaacaacatg caggaggcacccacagaata tgcatccatt tgtgtgagaa 300 gttaaacctg ccactgagcc aggcagcctacactgcatga gtgcctgtca atacctctgt 360 ctggaccttc agtttcaaat aaccttcaacctggaaagtt tcaattaaga tgctctgtgc 420 tggtgctgcg tcttaaaggt ccatgaagtatttagttaaa atttctcctg aaaactttgg 480 gagagttttg tacaagacag actcttccctgggaaaactc atattacgaa tagcagaata 540 atagggcttt taaaattgac catgtcgcaccatgtcgtac ccaaagggcg aattctgcag 600 atcagatct 609 109 1276 DNA Musmusculus 109 agatctctag ggaggaagag gaagtttgcc cttacctgac acgtgctgggaatgaagaca 60 gtgcctgcca tgcttgggac tcctcggtta tttagggaat tcttcatcctccatctgggc 120 ctctggagca tcctttgtga gaaagctact aagaggaatg atgaagagtgtccagtgcaa 180 cttactatta cgaggaattc caaacagtct gccaggacag gagagttatttaaaattcaa 240 tgtcctgtga aatactgtgt tcatagacct aatgtgactt ggtgtaagcacaatggaaca 300 atctgtgtac cccttgaggt tagccctcag ctatacacta gttgggaagaaaatcaatca 360 gttccggttt ttgttctcca ctttaaacca atacatctca gtgataatgggtcgtatagc 420 tgttctacaa acttcaattc tcaagttatt aatagccatt cagtaaccatccatgtgaca 480 gaaaggactc aaaactcttc agaacaccca ctaataacag tatctgacatcccagatgcc 540 accaatgcct caggaccatc caccatggaa gagaggccag gcaggacttggctgctttac 600 accttgcttc ctttgggggc attgcttctg ctccttgcct gtgtctgcctgctctgcttt 660 ctgaaaagga tccaagggaa agaaaagaag ccttctgact tggcaggaagggacactaac 720 ctggttgata ttccagccag ttccaggaca aatcaccaag cactgccatcaggaactgga 780 atttatgata atgatccctg gtctagcatg caggatgaat ctgaattgacaattagcttg 840 caatcagaga gaaacaacca gggcattgtt tatgcttctt tgaaccattgtgttattgga 900 aggaatccaa gacaggaaaa caacatgcag gaggcaccca cagaatatgcatccatttgt 960 gtgagaagtt aaacctgcca ctgagccagg cagcctacac tgcatgagtgcctgtcaata 1020 cctctgtctg gaccttcagt ttcaaataac cttcaacctg gaaagtttcaattaagatgc 1080 tctgtgctgg tgctgcgtct taaaggtcca tgaagtattt agttaaaatttctcctgaaa 1140 actttgggag agttttgtac aagacagact cttccctggg aaaactcatattacgaatag 1200 cagaataata gggcttttaa aattgaccat gtcgcaccat gtcgtacccaaagggcgaat 1260 tctgcagatc agatct 1276 110 486 DNA Mus musculus 110ggatcttgga gaaagctact aagaggaatg gagaaagcta ctaagaggaa tgatgaagag 60tgtccagtgc aacttactat tacgaggaat tccaaacagt ctgccaggac aggagagtta 120tttaaaattc aatgtcctgt gaaatactgt gttcatagac ctaatgtgac ttggtgtaag 180cacaatggaa caatctgtgt accccttgag gttagccctc agctatacac tagttgggaa 240gaaaatcaat cagttccggt ttttgttctc cactttaaac caatacatct cagtgataat 300gggtcgtata gctgttctac aaacttcaat tctcaagtta ttaatagcca ttcagtaacc 360atccatgtga cagaaaggac tcaaaactct tcagaacacc cactaataac agtatctgac 420atcccagatg ccaccaatgc ctcaggacca tccaccatgg aagagaggcc aggcaggact 480agatcc 486 111 464 DNA Mus musculus 111 agatcttgga gaaagctact aagaggaatgatgaagagtg tccagtgcaa cttactatta 60 cgaggaattc caaacagtct gccaggacaggagagttatt taaaattcaa tgtcctgtga 120 aatactgtgt tcatagacct aatgtgacttggtgtaagca caatggaaca atctgtgtac 180 cccttgaggt tagccctcag ctatacactagttgggaaga aaatcaatca gttccggttt 240 ttgttctcca ctttaaacca atacatctcagtgataatgg gtcgtatagc tgttctacaa 300 acttcaattc tcaagttatt aatagccattcagtaaccat ccatgtgaca gaaaggactc 360 aaaactcttc agaacaccca ctaataacagtatctgacat cccagatgcc accaatgcct 420 caggaccatc caccatggaa gagaggccaggcaggactag atct 464 112 538 DNA Mus musculus 112 atgaagacag tgcctgccatgcttgggact cctcggttat ttagggaatt cttcatcctc 60 catctgggcc tctggagcatcctttgtgag aaagctacta agaggaatga tgaagagtgt 120 ccagtgcaac ttactattacgaggaattcc aaacagtctc caggacagga gagttattta 180 aaattcaatg tcctgtgaaatactgtgttc atagacctaa tgtgacttgg tgtaagcaca 240 atggaacaat ctgtgtaccccttgaggtta gccctcagct atacactagt tgggaagaaa 300 atcaatcagt tccggtttttgttctccact ttaaaccaat acatctcagt gataatgggt 360 cgtatagctg ttctacaaacttcaattctc aagttattaa tagccattca gtaaccatcc 420 atgtgacaga aaggactcaaaactcttcag aacacccact aataacagta tctgacatcc 480 cagatgccac caatgcctcaggaccatcca ccatggaaga gaggccaggc aggactgc 538 113 537 DNA Mus musculus113 atgaagacag tgcctgccat gcttgggact cctcggttat ttagggaatt cttcatcctc 60catctgggcc tctggagcat cctttgtgag aaagctacta agaggaatga tgaagagtgt 120gaagtgcaac ttaatattaa gaggaattcc aaacactctg cctggacagg agagttattt 180aaaattgaat gtcctgtgaa atactgtgtt catagacctc atgtgacttg gtgtaagcac 240aatggaacaa tctgggtacc ccttgaagtt ggtcctcagc tatacactag ttgggaagaa 300aatcgatcag ttccggtttt tgttctccat tttaaaccaa tacatctcag tgataacggg 360tcgtatagct gttctacaaa cttcaattct caagttatta atagccattc agtaaccatc 420catgtgagag aaaggactca aaactcttca gaacacccac taataacagt atctgacatc 480ccagatgcca ccaatgcctc aggaccatcc accatggaag agaggccagg caggact 537

What is claimed is: 1) A purified polypeptide comprising an amino acidsequence set forth in SEQ ID NO:3. 2) The polypeptide of claim 1,wherein the amino acid sequence comprises an immunoglobulin like domain.3) The polypeptide of claim 1, wherein the polypeptide does not includea transmembrane domain. 4) The polypeptide of claim 1, wherein thepolypeptide does not include SEQ ID NO:5. 5) The polypeptide of claim 1,wherein the polypeptide does not include a tyrosine based domain. 6) Thepolypeptide of claim 1, wherein the polypeptide does not include SEQ IDNO:7. 7) The polypeptide of claim 1, wherein the polypeptide does notinclude SEQ ID NO:9. 8) The polypeptide of claim 1, wherein thepolypeptide does not include SEQ ID NO:
 11. 9) The polypeptide of claim1, wherein the polypeptide includes SEQ ID NO: 12 or SEQ ID NO:
 13. 10)The polypeptide of claim 1, wherein the polypeptide includes atransmembrane domain. 11) The polypeptide of claim 1, wherein thepolypeptide includes SEQ ID NO:14 or SEQ ID NO:15. 12) The polypeptideof claim 1, wherein the polypeptide includes a tyrosine based domain.13) The polypeptide of claim 1, wherein the polypeptide includes SEQ IDNO: 19, SEQ ID NO:20, or SEQ ID NO:21. 14) The polypeptide of claim 1,further comprising a second amino acid sequence operably linked to thefirst amino acid sequence. 15) The polypeptide of claim 1, furthercomprising a second amino acid sequence, wherein the second sequence ismore soluble in aqueous solution than the first amino acid sequence. 16)The polypeptide of claim 1, further comprising a detectable marker. 17)The polypeptide of claim 1, further comprising an affinity tag. 18) Thepolypeptide of claim 1, further comprising a second amino acid sequenceset forth in SEQ ID NO:97. 19) A purified polypeptide comprising anamino acid sequence set forth in SEQ ID NO:45. 20) The polypeptide ofclaim 19, wherein the amino acid sequence comprises an immunoglobulinlike domain. 21) The polypeptide of claim 19, wherein the polypeptidedoes not include a transmembrane domain. 22) The polypeptide of claim19, wherein the polypeptide does not include SEQ ID NO:47. 23) Thepolypeptide of claim 19, wherein the polypeptide does not include atyrosine based domain. 24) The polypeptide of claim 19, wherein thepolypeptide does not include SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:11.25) The polypeptide of claim 19, wherein the polypeptide includes SEQ IDNO:54 or SEQ ID NO:55. 26) The polypeptide of claim 19, wherein thepolypeptide includes a transmembrane domain. 27) The polypeptide ofclaim 19, wherein the polypeptide includes SEQ ID NO:56 or SEQ ID NO:57.28) The polypeptide of claim 19, wherein the polypeptide includes atyrosine based domain. 29) The polypeptide of claim 19, wherein thepolypeptide includes SEQ ID NO:61, SEQ ID NO:62, or SEQ ID NO:63. 30)The polypeptide of claim 19, further comprising a second amino acidsequence operably linked to the first amino acid sequence. 31) Thepolypeptide of claim 19, further comprising a second amino acidsequence, wherein the second sequence is more soluble in aqueoussolution than the first amino acid sequence. 32) The polypeptide ofclaim 19, further comprising a detectable marker. 33) The polypeptide ofclaim 19, further comprising an affinity tag. 34) The polypeptide ofclaim 19, further comprising a second amino acid sequence set forth inSEQ ID NO:97. 35) A purified polypeptide comprising an amino acidsequence set forth in SEQ ID NO:88. 36) The polypeptide of claim 35,wherein the amino acid sequence comprises an immunoglobulin like domain.37) The polypeptide of claim 35, wherein the polypeptide does notinclude a transmembrane domain. 38) The polypeptide of claim 35, whereinthe polypeptide does not include a tyrosine based domain. 39) Thepolypeptide of claim 35, wherein the polypeptide does not include SEQ IDNO:7, SEQ ID NO:9, or SEQ ID NO:
 11. 40) The polypeptide of claim 35,wherein the polypeptide includes a transmembrane domain. 41) Thepolypeptide of claim 35, wherein the polypeptide includes a tyrosinebased domain. 42) The polypeptide of claim 35, further comprising asecond amino acid sequence operably linked to the first amino acidsequence. 43) The polypeptide of claim 35, further comprising a secondamino acid sequence, wherein the second sequence is more soluble inaqueous solution than the first amino acid sequence. 44) The polypeptideof claim 35, further comprising a detectable marker. 45) The polypeptideof claim 35, further comprising an affinity tag. 46) The polypeptide ofclaim 35, further comprising a second amino acid sequence set forth inSEQ ID NO:97. 47) A purified polypeptide, comprising an amino acidsequence set forth in SEQ ID NO:7. 48) The polypeptide of claim 47,wherein the amino acid sequence comprises a tyrosine based domain. 49)The polypeptide of claim 47, wherein the polypeptide further includesSEQ ID NO:9. 50) The polypeptide of claim 47, wherein the polypeptidefurther includes SEQ ID NO:
 11. 51) The polypeptide of claim 47, whereinthe polypeptide further includes SEQ ID NO:9 and SEQ ID NO:
 11. 52) Thepolypeptide of claim 47, wherein the polypeptide does not include atransmembrane domain. 53) The polypeptide of claim 47, wherein thepolypeptide does not include SEQ ID NO:5 or SEQ ID NO:47. 54) Thepolypeptide of claim 47, wherein the polypeptide does not include animmunoglobulin like domain. 55) The polypeptide of claim 47, wherein thepolypeptide does not include SEQ ID NO:3, SEQ ID NO:45, or SEQ ID NO:88.56) The polypeptide of claim 47, wherein the polypeptide includes SEQ IDNO:
 16. 57) The polypeptide of claim 47, wherein the polypeptideincludes SEQ ID NO:
 17. 58) The polypeptide of claim 47, wherein thepolypeptide includes SEQ ID NO:
 18. 59) The polypeptide of claim 47,wherein the polypeptide includes SEQ ID NO:58. 60) The polypeptide ofclaim 47, wherein the polypeptide includes SEQ ID NO:59. 61) Thepolypeptide of claim 47, wherein the polypeptide includes SEQ ID NO:60.62) The polypeptide of claim 47, wherein the tyrosine based domaincomprises a phosphorylation site. 63) The polypeptide of claim 47,further comprising a second amino acid sequence operably linked to thefirst amino acid sequence. 64) The polypeptide of claim 47, furthercomprising a detectable marker. 65) The polypeptide of claim 47, furthercomprising an affinity tag. 66) A purified polypeptide, comprising anamino acid sequence set forth in SEQ ID NO:9. 67) The polypeptide ofclaim 66, wherein the amino acid sequence comprises a tyrosine baseddomain. 68) The polypeptide of claim 66, wherein the polypeptide furtherincludes SEQ ID NO:7. 69) The polypeptide of claim 66, wherein thepolypeptide further includes SEQ ID NO:
 11. 70) The polypeptide of claim66, wherein the polypeptide further includes SEQ ID NO:7 and SEQ ID NO:11. 71) The polypeptide of claim 66, wherein the polypeptide does notinclude a transmembrane domain. 72) The polypeptide of claim 66, whereinthe polypeptide does not include SEQ ID NO:5 or SEQ ID NO:47. 73) Thepolypeptide of claim 66, wherein the polypeptide does not include animmunoglobulin like domain. 74) The polypeptide of claim 66, wherein thepolypeptide does not include SEQ ID NO:3, SEQ ID NO:45, or SEQ ID NO:88.75) The polypeptide of claim 66, wherein the polypeptide includes SEQ IDNO:16. 76) The polypeptide of claim 66, wherein the polypeptide includesSEQ ID NO:
 17. 77) The polypeptide of claim 66, wherein the polypeptideincludes SEQ ID NO:
 18. 78) The polypeptide of claim 66, wherein thepolypeptide includes SEQ ID NO:58. 79) The polypeptide of claim 66,wherein the polypeptide includes SEQ ID NO:59. 80) The polypeptide ofclaim 66, wherein the polypeptide includes SEQ ID NO:60. 81) Thepolypeptide of claim 66, wherein the tyrosine based domain comprises aphosphorylation site. 82) The polypeptide of claim 66, furthercomprising a second amino acid sequence operably linked to the firstamino acid sequence. 83) The polypeptide of claim 66, further comprisinga detectable marker. 84) The polypeptide of claim 66, further comprisingan affinity tag. 85) A purified polypeptide, comprising an amino acidsequence set forth in SEQ ID NO:
 11. 86) The polypeptide of claim 85,wherein the amino acid sequence comprises a tyrosine based domain. 87)The polypeptide of claim 85, wherein the polypeptide further includesSEQ ID NO:7. 88) The polypeptide of claim 85, wherein the polypeptidefurther includes SEQ ID NO:9. 89) The polypeptide of claim 85, whereinthe polypeptide further includes SEQ ID NO:7 and SEQ ID NO:9. 90) Thepolypeptide of claim 85, wherein the polypeptide does not include atransmembrane domain. 91) The polypeptide of claim 85, wherein thepolypeptide does not include SEQ ID NO:5. 92) The polypeptide of claim85, wherein the polypeptide does not include SEQ ID NO:47. 93) Thepolypeptide of claim 85, wherein the polypeptide does not include animmunoglobulin like domain. 94) The polypeptide of claim 85, wherein thepolypeptide does not include SEQ ID NO:3 or SEQ ID NO:45. 95) Thepolypeptide of claim 85, wherein the polypeptide includes SEQ ID NO: 16.96) The polypeptide of claim 85, wherein the polypeptide includes SEQ IDNO:
 17. 97) The polypeptide of claim 85, wherein the polypeptideincludes SEQ ID NO:
 18. 98) The polypeptide of claim 85, wherein thepolypeptide includes SEQ ID NO:58. 99) The polypeptide of claim 85,wherein the polypeptide includes SEQ ID NO:59. 100) The polypeptide ofclaim 85, wherein the polypeptide includes SEQ ID NO:60. 101) Thepolypeptide of claim 85, wherein the tyrosine based domain comprises aphosphorylation site. 102) The polypeptide of claim 85, furthercomprising a second amino acid sequence operably linked to the firstamino acid sequence. 103) The polypeptide of claim 85, furthercomprising a detectable marker. 104) The polypeptide of claim 85,further comprising an affinity tag. 105) A purified polypeptidecomprising an amino acid sequence that is 95% or more identical to SEQID NO:3 and includes an immunoglobulin like domain structure. 106) Thepolypeptide of claim 105, wherein the immunoglobulin like domainstructure is substantially similar to the structure of SEQ ID NO:3. 107)The polypeptide of claim 105, wherein the immunoglobulin like domainstructure is substantially similar to the structure of SEQ ID NO: 12.108) The polypeptide of claim 105, wherein the amino acid sequence is98% or more identical to SEQ ID NO:3. 109) The polypeptide of claim 105,wherein the polypeptide does not include SEQ ID NO:5. 110) Thepolypeptide of claim 105 wherein the polypeptide does not include SEQ IDNO:7, SEQ ID NO:9, or SEQ ID NO:11. 111) The polypeptide of claim 105,wherein the polypeptide further includes SEQ ID NO:7, SEQ ID NO:9, orSEQ ID NO:11. 112) The polypeptide of claim 105, further comprising asecond amino acid sequence operably linked with the first amino acidsequence. 113) The polypeptide of claim 112, wherein the second aminoacid sequence includes a segment that is 95% or more identical to SEQ IDNO:16. 114) The polypeptide of claim 113, wherein the segment includes apolymer set forth in SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:
 11. 115)The polypeptide of claim 105, wherein the polypeptide is operably linkedto a detectable marker or an affinity tag. 116) A purified polypeptidecomprising an amino acid sequence that is 95% or more identical to SEQID NO:45 and includes an immunoglobulin like domain structure. 117) Thepolypeptide of claim 116, wherein the immunoglobulin like domainstructure is substantially similar to the structure of SEQ ID NO:45.118) The polypeptide of claim 116, wherein the immunoglobulin likedomain structure is substantially similar to the structure of SEQ IDNO:54. 119) The polypeptide of claim 116, wherein the amino acidsequence is 98% or more identical to SEQ ID NO:45. 120) The polypeptideof claim 116, wherein the polypeptide does not include SEQ ID NO:47.121) The polypeptide of claim 116, wherein the polypeptide does notinclude SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:
 11. 122) The polypeptideof claim 116, wherein the polypeptide further includes SEQ ID NO:7, SEQID NO:9, or SEQ ID NO:11. 123) The polypeptide of claim 116, furthercomprising a second amino acid sequence operably linked with the firstamino acid sequence. 124) The polypeptide of claim 123, wherein thesecond amino acid sequence includes a segment that is 95% or moreidentical to SEQ ID NO:58. 125) The polypeptide of claim 124, whereinthe segment includes a polymer set forth in SEQ ID NO:7, SEQ ID NO:9, orSEQ ID NO:
 11. 126) The polypeptide of claim 116, wherein thepolypeptide is operably linked to a detectable marker or an affinitytag. 127) A purified polypeptide comprising an amino acid sequence thatis 95% or more identical to SEQ ID NO:88 and includes an immunoglobulinlike domain structure. 128) The polypeptide of claim 127, wherein theimmunoglobulin like domain structure is substantially similar to thestructure of SEQ ID NO:88. 129) The polypeptide of claim 127, whereinthe immunoglobulin like domain structure is substantially similar to thestructure of SEQ ID NO:86. 130) The polypeptide of claim 127, whereinthe amino acid sequence is 98% or more identical to SEQ ID NO:88. 131)The polypeptide of claim 127, wherein the polypeptide further includesSEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:
 11. 132) The polypeptide ofclaim 127, further comprising a second amino acid sequence operablylinked with the first amino acid sequence. 133) The polypeptide of claim132, wherein the second amino acid sequence comprises a detectablemarker or an affinity tag. 134) The polypeptide of claim 127, whereinthe polypeptide is operably linked to a detectable marker or an affinitytag. 135) A purified immunogenic polypeptide, comprising ten consecutiveamino acid residues of SEQ ID NO:3. 136) A purified immunogenicpolypeptide, consisting essentially of 8 to 40 consecutive amino acidresidues of SEQ ID NO:3. 137) A purified polynucleotide, comprising anucleic acid sequence encoding a SPEX polypeptide, or a complement ofthe nucleic acid sequence. 138) The polynucleotide of claim 137, whereinthe polynucleotide includes no more than 30000 nucleotides or nucleotidebase pairs. 139) The polynucleotide of claim 137, wherein thepolynucleotide includes no more than 15000 nucleotides or nucleotidebase pairs. 140) The polynucleotide of claim 137, wherein thepolynucleotide includes no more than 7000 nucleotides or nucleotide basepairs. 141) The polynucleotide of claim 137, wherein the polynucleotideincludes 800 to 5000 nucleotides or nucleotide base pairs. 142) Thepolynucleotide of claim 137, wherein the polynucleotide includes anintervening sequence. 143) The polynucleotide of claim 137, wherein thepolypeptide does not include a transmembrane domain. 144) Thepolynucleotide of claim 137, wherein the polypeptide does not includeSEQ ID NO:5. 145) The polynucleotide of claim 137, wherein thepolypeptide does not include SEQ ID NO:7. 146) The polynucleotide ofclaim 137, wherein the polypeptide does not include SEQ ID NO:9. 147)The polynucleotide of claim 137, wherein the polypeptide does notinclude SEQ ID NO:11. 148) The polynucleotide of claim 137, wherein thepolypeptide includes SEQ ID NO:3. 149) The polynucleotide of claim 137,wherein the polypeptide includes SEQ ID NO:
 12. 150) The polynucleotideof claim 137, wherein the polypeptide includes SEQ ID NO:
 13. 151) Thepolynucleotide of claim 137, wherein the polypeptide includes SEQ IDNO:14. 152) The polynucleotide of claim 137, wherein the polypeptideincludes SEQ ID NO:
 15. 153) The polynucleotide of claim 137, whereinthe polypeptide includes SEQ ID NO:19. 154) The polynucleotide of claim137, wherein the polypeptide includes SEQ ID NO:20. 155) Thepolynucleotide of claim 137, wherein the polypeptide includes SEQ IDNO:21. 156) A purified polynucleotide, comprising a nucleic acidsequence encoding a polypeptide sequence set forth in SEQ ID NO:45, or acomplement of the nucleic acid sequence. 157) The polynucleotide ofclaim 156, wherein the polynucleotide includes no more than 30000nucleotides or nucleotide base pairs. 158) The polynucleotide of claim156, wherein the polynucleotide includes no more than 15000 nucleotidesor nucleotide base pairs. 159) The polynucleotide of claim 156, whereinthe polynucleotide includes no more than 7000 nucleotides or nucleotidebase pairs. 160) The polynucleotide of claim 156, wherein thepolynucleotide includes 800 to 5000 nucleotides or nucleotide basepairs. 161) The polynucleotide of claim 156, wherein the polynucleotideincludes an intervening sequence. 162) The polynucleotide of claim 156,wherein the polypeptide does not include a transmembrane domain. 163)The polynucleotide of claim 156, wherein the polypeptide does notinclude SEQ ID NO:47. 164) The polynucleotide of claim 156, wherein thepolypeptide does not include SEQ ID NO:7. 165) The polynucleotide ofclaim 156, wherein the polypeptide does not include SEQ ID NO:9. 166)The polynucleotide of claim 156, wherein the polypeptide does notinclude SEQ ID NO:11. 167) The polynucleotide of claim 156, wherein thepolypeptide includes SEQ ID NO:54. 168) The polynucleotide of claim 156,wherein the polypeptide includes SEQ ID NO:55. 169) The polynucleotideof claim 156, wherein the polypeptide includes SEQ ID NO:56. 170) Thepolynucleotide of claim 156, wherein the polypeptide includes SEQ IDNO:57. 171) The polynucleotide of claim 156, wherein the polypeptideincludes SEQ ID NO:61. 172) The polynucleotide of claim 156, wherein thepolypeptide includes SEQ ID NO:62. 173) The polynucleotide of claim 156,wherein the polypeptide includes SEQ ID NO:63. 174) A purifiedpolynucleotide, comprising a nucleic acid sequence encoding apolypeptide sequence set forth in SEQ ID NO:88, or a complement of thenucleic acid sequence. 175) The polynucleotide of claim 174, wherein thepolynucleotide includes no more than 30000 nucleotides or nucleotidebase pairs. 176) The polynucleotide of claim 174, wherein thepolynucleotide includes no more than 15000 nucleotides or nucleotidebase pairs. 177) The polynucleotide of claim 174, wherein thepolynucleotide includes no more than 7000 nucleotides or nucleotide basepairs. 178) The polynucleotide of claim 174, wherein the polynucleotideincludes 800 to 5000 nucleotides or nucleotide base pairs. 179) Thepolynucleotide of claim 174, wherein the polynucleotide includes anintervening sequence. 180) The polynucleotide of claim 174, wherein thepolypeptide does not include a transmembrane domain. 181) Thepolynucleotide of claim 174, wherein the polypeptide does not includeSEQ ID NO:7. 182) The polynucleotide of claim 174, wherein thepolypeptide does not include SEQ ID NO:9. 183) The polynucleotide ofclaim 174, wherein the polypeptide does not include SEQ ID NO:
 11. 184)The polynucleotide of claim 174, wherein the polypeptide includes SEQ IDNO:85. 185) The polynucleotide of claim 174, wherein the polypeptideincludes SEQ ID NO:86. 186) A purified polynucleotide, comprising anucleic acid sequence encoding a SPEX tyrosine based domain, or acomplement of the nucleic acid sequence. 187) A purified polynucleotide,comprising: a) a nucleic acid sequence encoding a SPEX polypeptide, or acomplement thereof; and b) a vector sequence operatively linked to thenucleic acid sequence. 188) The polynucleotide of claim 187, wherein thevector sequence comprises an expression vector having one or morecontrol elements for producing an expression product of the nucleic acidsequence. 189) The polynucleotide of claim 188, wherein the expressionvector comprises an in vitro expression system. 190) The polynucleotideof claim 188, wherein the expression vector comprises a cellularexpression system. 191) The expression vector of 190, wherein theexpression vector includes one or more control elements for expressingthe product in a human cell. 192) The polynucleotide of claim 190,wherein the expression vector includes one or more control elements forexpressing the product in one or more cell types selected from a groupconsisting of: a bacterial cell, a yeast cell, an insect cell, and amammalian cell. 193) The polynucleotide of claim 192, wherein theproduct comprises the complement of the nucleic acid sequence. 194) Thepolynucleotide of claim 192, wherein the product comprises the SPEXpolypeptide. 195) The polypeptide of claim 194, wherein the polypeptideincludes SEQ ID NO:3. 196) The polypeptide of claim 195, wherein thepolypeptide does not include a transmembrane domain. 197) Thepolypeptide of claim 194, wherein the polypeptide includes SEQ ID NO:45.198) The polypeptide of claim 197, wherein the polypeptide does notinclude a transmembrane domain. 199) The polypeptide of claim 194,wherein the polypeptide includes SEQ ID NO:88. 200) The polypeptide ofclaim 199, wherein the polypeptide does not include a transmembranedomain. 201) A purified SPEX expression vector, comprising: a nucleicacid sequence encoding a SPEX polypeptide operably linked to a vectorsequence having one or more control elements for expressing the SPEXpolypeptide. 202) The SPEX expression vector of claim 201, wherein theSPEX polypeptide comprises a fusion of a first SPEX amino acid sequenceoperably linked with a second amino acid sequence. 203) The SPEXexpression vector of claim 202, wherein the second amino acid sequencecomprises a solubility enhancing polypeptide. 204) The SPEX expressionvector of claim 202, wherein the solubility enhancing polypeptidecomprises an Ig domain from IgG. 205) The SPEX expression vector ofclaim 202, wherein the solubility enhancing polypeptide comprises SEQ IDNO:97. 206) The SPEX expression vector of claim 205, wherein the SPEXpolypeptide comprises SEQ ID NO:3. 207) of claim 202, wherein the secondamino acid sequence comprises a tag. 208) The polynucleotide of claim187, comprising pEYFP-N1. 209) The polynucleotide of claim 187, whereinthe polynucleotide comprises 210) A purified host cell, comprising aSPEX expression vector including a nucleic acid sequence encoding a SPEXexpression product operably linked with a vector sequence having one ormore control elements for expressing the SPEX expression product. 211)The host cell of claim 210, wherein the SPEX expression productcomprises a polynucleotide that is complementary to a nucleic acidpolymer encoding a SPEX polypeptide. 212) The host cell of claim 210,wherein the SPEX expression product comprises a SPEX polypeptide. 213)The host cell of claim 210, comprising a bacterial cell. 214) The hostcell of claim 210, comprising a yeast cell. 215) The host cell of claim210, comprising an insect cell. 216) The host cell of claim 210,comprising a mammalian cell. 217) The host cell of claim 210, comprisinga human cell. 218) An isolated antibody that immunoreacts with a SPEXpolypeptide. 219) The antibody of claim 218, wherein the antibodycomprises a monoclonal antibody, or immunoreactive fragment thereof.220) The antibody of claim 218, wherein the antibody comprises a humanmonoclonal antibody, a humanized monoclonal antibody, or immunoreactivefragment thereof. 221) The antibody of claim 218, wherein the SPEXpolypeptide comprises a human SPEX polypeptide. 222) The antibody ofclaim 218, wherein the antibody immunoreacts with an extracellulardomain of a SPEX polypeptide. 223) The antibody of claim 222, whereinthe antibody comprises a monoclonal antibody, or immunoreactive fragmentthereof. 224) The antibody of claim 223, wherein the antibody modulatesa metabolism of a cell expressing the SPEX polypeptide. 225) Theantibody of claim 224, wherein the cell comprises a lymphocyte. 226) Theantibody of claim 225, wherein the metabolism comprises a cellularproliferation. 227) The antibody of claim 226, wherein the cellularproliferation is stimulated. 228) The antibody of claim 226, wherein thecellular proliferation is inhibited. 229) The antibody of claim 218,wherein the antibody immunoreacts with an extracellular domain of a SPEXpolypeptide, but does not immunoreact with an intracellular domain of aSPEX polypeptide. 230) The antibody of claim 218, wherein the antibodyimmunoreacts with an intracellular domain of a SPEX polypeptide. 231)The antibody of claim 218, wherein the antibody immunoreacts with anintracellular domain of a SPEX polypeptide, but does not immunoreactwith an extracellular domain of a SPEX polypeptide. 232) The antibody ofclaim 218, wherein the antibody immunoreacts with an amino acid sequenceset forth in SEQ ID NO:21, SEQ ID NO:63, or SEQ ID NO:85. 233) Theantibody of claim 218, wherein the antibody immunoreacts with an aminoacid sequence set forth in SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO: 11.234) A method of identifying a SPEX binding partner, comprising: a)contacting a candidate binding partner with a SPEX polypeptide; b)detecting whether or not a binding complex is formed that includes theSPEX polypeptide; c) selecting the candidate binding partner as a SPEXbinding partner when the binding complex is formed. 235) The method ofclaim 234, wherein the SPEX polypeptide comprises SEQ ID NO:3, SEQ IDNO:45, or SEQ ID NO:88. 236) The method of claim 234, wherein the SPEXpolypeptide comprises an extracellular domain of the SPEX polypeptide.237) The method of claim 234, wherein the SPEX polypeptide comprises afusion polypeptide having a first amino acid including a SPEX Ig likedomain operably linked to a second amino acid including a constantregion of an IgG polypeptide. 238) The method of claim 234, wherein theSPEX polypeptide includes a detectable label. 239) The method of claim234, wherein the SPEX polypeptide comprises an intracellular domain ofthe SPEX polypeptide. 240) The method of claim 239, wherein the SPEXpolypeptide includes one or more of SEQ ID NO:7, SEQ ID NO:9, or SEQ IDNO:11. 241) The method of claim 234, wherein the candidate bindingpartner comprises a polypeptide. 242) The method of claim 234, whereinthe candidate binding partner comprises an organic chemical compound.243) A method of isolating a SPEX polypeptide from a biological samplecontaining the SPEX polypeptide, comprising: a) contacting thebiological sample with an affinity matrix having an anti-SPEX antibodybound to the matrix, to produce a complex including the SPEX polypeptideand the anti-SPEX antibody bound to the matrix; b) separating the matrixand the remainder of the biological sample; c) separating the SPEXpolypeptide and the anti-SPEX antibody; and d) collecting the isolatedSPEX polypeptide. 244) A method of modulating a metabolism of alymphocyte that expresses a SPEX polypeptide, comprising contacting thelymphocyte with an anti-SPEX antibody that immunoreacts with anextracellular domain of the SPEX polypeptide. 245) The method of claim244, wherein the metabolism comprises a proliferation. 246) The methodof claim 245, wherein the proliferation is stimulated. 247) The methodof claim 244, wherein the anti-SPEX antibody immunoreacts with animmunoglobulin like domain of the SPEX polypeptide. 248) The method ofclaim 244, wherein the anti-SPEX antibody immunoreacts with SEQ IDNO:13, SEQ ID NO:55, or SEQ ID NO:85. 249) The method of claim 244,wherein the antibody is a monoclonal antibody or a fragment thereofcapable of immunoreacting with an extracellular domain of a SPEXpolypeptide. 250) The method of claim 244, wherein the lymphocytes arein an in vitro culture.